Benzoxazinone derivatives for use in the treatment of angiogenesis

ABSTRACT

This invention relates to novel compounds of Formula (I) for use as vascular damaging agents: wherein: X is selected from: —O—, —S—, —S(O)—, —S(O 2 )—, —N(R 4 )— or —N(R 4 )CH 2 C(O)—; R 1  is independently selected from: amino, halo, hydroxy, —OPO 3 H 2 , C 1-4 alkyl, C 1-4 alkoxy, N—C 1-4 alkylamino, N,N-di-C 1-4 alkylamino, C 1-4 alkanoylamino or C 1-4 alkylthio wherein the amino group is optionally substituted by an amino acid residue and the hydroxy group is optionally esterified; R 2  is selected from: hydrogen or C 1-4 alkyl; R 3  is selected from: hydrogen or C 1-4 alkyl; R 4  is selected from: hydrogen or C 1-4 alkyl; n is 0, 1 or 2; and p is 0, 1, 2 or 3; or a salt, pro-drug or solvate thereof. The invention also relates to methods for preparing compounds of Formula (I), to their use as medicaments (including methods for the treatment of angiogenesis or disease states associated with angiogenesis) and to pharmaceutical compositions containing compounds of Formula (I).

[0001] This invention relates to vascular damaging agents and theiruses. In particular it relates to certain compounds which may be of useas vascular damaging agents, to methods for preparing the compounds, totheir use as medicaments (including methods for the treatment ofangiogenesis or disease states associated with angiogenesis) and topharmaceutical compositions containing them. The invention also relatesto the use of such compounds in the manufacture of medicaments for theproduction of anti-angiogenic and/or anti-vascular effects.

[0002] Normal angiogenesis plays an important role in a variety ofprocesses including embryonic development, wound healing and severalcomponents of female reproductive function. Undesirable or pathologicalangiogenesis has been associated with disease states including diabeticretinopathy, psoriasis, cancer, rheumatoid arthritis, atheroma, Kaposi'ssarcoma and haemangioma (Fan et al, 1995, Trends Pharmacol. Sci. 16:57-66; Folkman, 1995, Nature Medicine 1: 27-31). Formation of newvasculature by angiogenesis is a key pathological feature of severaldiseases (J. Folkman, New England Journal of Medicine 333, 1757-1763(1995)). For example, for a solid tumour to grow it must develop its ownblood supply upon which it depends critically for the provision ofoxygen and nutrients; if this blood supply is mechanically shut off thetumour undergoes necrotic death. Neovascularisation is also a clinicalfeature of skin lesions in psoriasis, of the invasive pannus in thejoints of rheumatoid arthritis patients and of atherosclerotic plaques.Retinal neovascularisation is pathological in macular degeneration andin diabetic retinopathy.

[0003] Reversal of neovascularisation by damaging the newly-formedvascular endothelium is therefore expected to have a beneficialtherapeutic effect. Such vascular-damaging activity would clearly be ofvalue in the treatment of disease states associated with angiogenesissuch as cancer, diabetes, psoriasis, rheumatoid arthritis, Kaposi'ssarcoma, haemangioma, acute and chronic nephropathies, atheroma,arterial restenosis, autoimmune diseases, acute inflammation,endometriosis, dysfunctional uterine bleeding and ocular diseases withretinal vessel proliferation.

[0004] Certain known compounds that cause selective destruction oftumour vasculature have been reported, in vitro and at non-cytotoxicconcentrations, to cause effects on proliferating endothelial cells, ie,cell detachment [Blakey D C et al, Proceedings of the AmericanAssociation for Cancer Research, 41, 329, 2000 abstract 2086] andchanges in cell shape [Davis P D et al, Proceedings of the AmericanAssociation for Cancer Research, 41, 329, 2000 abstract 2085; Chaplin DJ & Dougherty G J, Br J Cancer, 80, Suppl 1, 57-64, 1999]. It cantherefore be expected that these compounds will have damaging effects onnewly-formed vasculature, for example the vasculature of tumours. It canreasonably be predicted, for example, that they will be capable ofcausing selective destruction of tumour vasculature, both in vitro andin vivo. Destruction of tumour vasculature in turn leads to a reductionin tumour blood flow and to tumour cell death due to starvation ofoxygen and nutrients, ie, to anti-tumour activity [Davis P D et al;Chaplin D J & Dougherty G J; Blakey D C et al, all supra].

[0005] Compounds with this activity have also been described inInternational Patent Application WO 99/02166 (AngiogenePharmaceuticals), International Patent Application WO00/40529 (AngiogenePharmaceuticals) and International Patent Application WO 00/41669(Angiogene Pharmaceuticals).

[0006] We have identified a class of benzoxazinone compounds withtherapeutic activity. Thus, according to the first feature of thepresent invention there is provided the use of a compound of Formula (I)as a medicament, wherein:

[0007] X is selected from: —O—, —S—, —S(O)—, —S(O₂)—, —N(R₄)— or—N(R₄)CH₂C(O)—;

[0008] R₁ is independently selected from: amino, halo, hydroxy, —OPO₃H₂,C₁₋₄alkyl, C₁₋₄alkoxy, N—C₁₋₄alkylamino, N,N-di-C₁₋₄alkylamino,C₁₋₄alkanoylamino or C₁₋₄alkylthio wherein the amino group is optionallysubstituted by an amino acid residue and the hydroxy group is optionallyesterified;

[0009] R₂ is selected from: hydrogen or C₁₋₄alkyl;

[0010] R₃ is selected from: hydrogen or C₁₋₄alkyl;

[0011] R₄ is selected from: hydrogen or C₁₋₄alkyl;

[0012] n is 0, 1 or 2; and

[0013] p is 0, 1, 2 or 3;

[0014] or a pharmaceutically-acceptable salt, pro-drug or solvatethereof.

[0015] According to a further aspect of the first feature of theinvention there is provided a pharmaceutical composition comprising acompound of Formula (I) or pharmaceutically-acceptable salt, pro-drug orsolvate thereof, and a pharmaceutically-acceptable excipient.

[0016] According to a further aspect of the first feature of theinvention there is provided a pharmaceutical composition comprising acompound of Formula (I), or a pharmaceutically-acceptable salt, pro-drugor solvate thereof, in admixture with a pharmaceutically-acceptablediluent or carrier.

[0017] Compounds of Formula (I) have vascular damaging activity. Thus,according to the second feature of the present invention there isprovided the use of a compound of Formula (I) orpharmaceutically-acceptable salt, pro-drug or solvate thereof, for themanufacture of a medicament to inhibit and/or reverse and/or alleviatesymptoms of angiogenesis and/or any disease state associated withangiogenesis.

[0018] According to a further aspect of the second feature of theinvention there is provided a method of treatment, in a warm-bloodedanimal, to inhibit and/or reverse and/or alleviate symptoms ofangiogenesis and/or any disease state associated with angiogenesiscomprising administering to said warm-blooded animal a therapeutically(including prophylactically) effective amount of a compound of Formula(I), or a pharmaceutically-acceptable salt, pro-drug or solvate thereof.

[0019] Preferably a warm-blooded animal is a human.

[0020] The invention also provides a class of novel benzoxazinonecompounds. Thus, according to a third feature of the invention there isprovided a compound of Formula (II), wherein:

[0021] X is selected from: —O—, —S—, —S(O)—, —S(O₂), —N(R₄)— or—N(R₄)CH₂C(O)—;

[0022] R₁ is independently selected from: amino, halo, hydroxy, —OPO₃H₂,C₁₋₄alkyl, C₁₋₄alkoxy, N—C₁₋₄alkylamino, N,N-di-C₁₋₄alkylamino,C₁₋₄alkanoylamino or C₁₋₄alkylthio wherein the amino group is optionallysubstituted by an amino acid residue and the hydroxy group is optionallyesterified;

[0023] R₂ is selected from: hydrogen or C₁₋₄alkyl;

[0024] R₃ is selected from: hydrogen or C₁₋₄alkyl;

[0025] R₄ is selected from: hydrogen or C₁₋₄alkyl;

[0026] n is 0, 1 or 2; and

[0027] p is 0, 1, 2 or 3;

[0028] with the proviso that the following compounds are excluded:

[0029] 6-benzyloxy-2H-1,4-benzoxazin-3(4H)-one;

[0030] 7-benzyloxy-2H-1,4-benzoxazin-3(4H)-one;

[0031] 2-methyl-7-benzyloxy-2H-1,4-benzoxazin-3(4H)-one;

[0032] 2,4-dimethyl-7-benzyloxy-2H-1,4-benzoxazin-3(4H)-one;

[0033] 2-methyl-7-(3,5-dichlorobenzyloxy)-2H-1,4-benzoxazin-3(4H)-one;

[0034]2,4-dimethyl-7-(3,5-dichlorobenzyloxy)-2H-1,4-benzoxazin-3(4H)-one;

[0035] 7-phenylthio-2H-1,4-benzoxazin-3(4H)-one;

[0036] 4-methyl-7-phenylthio-2H-1,4-benzoxazin-3(4H)-one;

[0037] 4-methyl-7-phenylsulfinyl-2H-1,4-benzoxazin-3(4H)-one; and

[0038] 6-phenylsulfonyl-2H-1,4-benzoxazin-3(4H)-one.

[0039] or a salt, pro-drug or solvate thereof.

[0040] According to a further aspect of the third feature of theinvention there is provided a compound of Formula (II), or a salt,pro-drug or solvate thereof, as defined above, with the proviso that

[0041] (i) when the group —(CH₂)_(n)—X— is linked at the 6-position ofthe benzoxazinone ring, n is 0 or 1, X is —O— or —S(O₂)—, p is 0 and R₂is hydrogen then R₃ cannot be hydrogen; and

[0042] (ii) when the group —(CH₂)_(n)—X— is linked at the 7-position ofthe benzoxazinone ring, n is 0 or 1, X is —O—, —S— or —S(O)—, p is 0 or(R₁)_(p) is 3,5-dichloro, and R₂ is hydrogen or methyl then R₃ cannot behydrogen or methyl.

[0043] According to a further aspect of the third feature of theinvention there is provided a compound of Formula (IIa), wherein:

[0044] X is selected from: —O—, —S—, —S(O)—, —S(O₂)—, —N(R₄)— or—N(R₄)CH₂C(O)—;

[0045] R₁ is independently selected from: amino, hydroxy, —OPO₃H₂,C₁₋₄alkyl, C₁₋₄alkoxy, N—C₁₋₄alkylamino, N,N-di-C₁₋₄alkylamino,C₁₋₄alkanoylamino or C₁₋₄alkylthio wherein the amino group is optionallysubstituted by an amino acid residue and the hydroxy group is optionallyesterified;

[0046] R₂ is selected from: hydrogen or C₁₋₄alkyl;

[0047] R₃ is selected from: hydrogen or C₁₋₄alkyl;

[0048] R₄ is selected from: hydrogen or C₁₋₄alkyl;

[0049] n is 0, 1 or 2; and

[0050] p is 1, 2 or 3;

[0051] or a salt, pro-drug or solvate thereof.

[0052] According to a further aspect of the third feature of theinvention there is provided a pharmaceutical composition comprising acompound of Formula (II) or Formula (IIa) or pharmaceutically-acceptablesalt, pro-drug or solvate thereof, and a pharmaceutically-acceptableexcipient.

[0053] According to a further aspect of the third feature of theinvention there is provided a pharmaceutical composition comprising acompound of Formula (II), or Formula (IIa) or apharmaceutically-acceptable salt, pro-drug or solvate thereof, inadmixture with a pharmaceutically-acceptable diluent or carrier.

[0054] According to a further aspect of the third feature of the presentinvention there is provided the use of a compound of Formula (II) orFormula (IIa) or pharmaceutically-acceptable salt, pro-drug or solvatethereof, for the manufacture of a medicament to inhibit and/or reverseand/or alleviate symptoms of angiogenesis and/or any disease stateassociated with angiogenesis.

[0055] According to a further aspect of the third feature of theinvention there is provided a method of treatment, in a warm-bloodedanimal, to inhibit and/or reverse and/or alleviate symptoms ofangiogenesis and/or any disease state associated with angiogenesiscomprising administering to said warm-blooded animal a therapeutically(including prophylactically) effective amount of a compound of Formula(II) or Formula (IIa), or a pharmaceutically-acceptable salt, pro-drugor solvate thereof.

[0056] Whilst pharmaceutically-acceptable salts of compounds of theinvention are preferred, other non-pharmaceutically-acceptable salts ofcompounds of the invention may also be useful, for example in thepreparation of pharmaceutically-acceptable salts of compounds of theinvention.

[0057] For the avoidance of doubt when p is 0, all positions on thephenyl ring are substituted by hydrogen.

[0058] For the avoidance of doubt the use of the term (R₁)_(p) when p isbetween 1 and 3, means that there are 1, 2 or 3 R¹ substituents on thephenyl ring, which when p is 2 or 3 can be the same group or differentgroups. For example, where (R₁)_(p) is 3-chloro-4-methoxy then p is 2and the phenyl ring has a chloro group at the 3-position and a methoxygroup at the 4-position, in relation to the —(CH₂)_(n)X— group.

[0059] The term halo refers to fluoro, chloro, bromo or iodo.

[0060] The term carbamoyl refers to the group —CONH₂.

[0061] An amino acid residue is defined as that derived from thecoupling of an L-amino acid with an amino group via an amide bond. Thisbond can either be formed via a carboxylate group on the amino acidbackbone or via a side chain carboxylate group, preferably via acarboxylate group on the amino acid backbone. Amino acid residuesinclude those derived from natural and non-natural amino acids,preferably natural amino acids and include α-amino acids β-amino acidsand γ-amino acids. For the avoidance of doubt amino acids include thosewith the generic structure:

[0062] where R is the amino acid side chain. The definition of aminoacid also includes amino acid analogues which have additional methylenegroups within the amino acid backbone, for example β-alanine and aminoacids which are not naturally occurring such as cyclohexylalanine.

[0063] Preferred amino acids include glycine, alanine, valine, leucine,isoleucine, methionine, proline, phenylalanine, tryptophan, serine,threonine, cysteine, tyrosine, asparaginine, glutamine, aspartic acid,glutamic acid, lysine, arginine, histidine, β-alanine and ornithine.More preferred amino acids include glutamic acid, serine, threonine,arginine, glycine, alanine, β-alanine and lysine. Especially preferredamino acids include glutamic acid, serine, and glycine.

[0064] Esterifying groups at R₁ are esterifying groups which increasethe solubility of the molecule in water at a pH of approximately pH=7.Such groups include groups with ionisable groups, such as acidicfunctions or basic functions and groups containing a hydrophilicfunction. Basic functions include: amino, morpholino, piperidino,piperazino, pyrrolidino, amino acids and imidazolino. Acidic functionsinclude: carboxy, sulphonic acid, phosphate, sulphate and acid mimeticssuch as tetrazolyl. Hydrophilic groups include hydroxyl.

[0065] Suitable R₁ groups wherein hydroxy is esterfied include:C₁₋₆alkanoyloxy, arylcarbonyloxy, heterocyclylcarbonyloxy,heteroarylcarbonyloxy wherein the R₁ group is optionally substitutedwith between 1 and 3 groups selected from C₁₋₄alkyl, C₁₋₄alkanoyl,C₁₋₄alkanoylC₁₋₄alkyl, C₁₋₄alkanoylheterocyclyl, hydroxy,hydroxyC₁₋₄alkyl, carboxy, carboxyphenyl, phosphono, phosphonoC₁₋₄alkyl,amino, aminoC₁₋₄alkyl, N-C₁₋₄alkylamino, N,N-diC₁₋₄alkylamino,carbamoyl, carbamoyl₁₋₄alkyl, heterocyclyl, heterocyclylC₁₋₄alkyl,heterocyclylcarbonyl, heterocyclC₁₋₄alkanoylamino,carbamoylheterocyclyl, [wherein optional substituents comprisingheterocyclyl are optionally further substituted by C₁₋₄alkyl,hydroxyC₁₋₄alkyl, C₁₋₄alkoxyC₁₋₄alkyl, C₁₋₄alkanoyl and formyl, whereinthe carbamoyl and amino optional substituents are optionally furtherN-substituted by C₁₋₄alkyl, di-C₁₋₄alkyl, hydroxyC₁₋₄alkyl,di-(hydroxyC₁₋₄alkyl), carboxyC₁₋₄alkyl, and wherein the amino group isoptionally substituted by an amino acid residue] with the proviso thatwhen R₁ is C₁₋₆alkanoyloxy or arylcarbonyloxy R₁ is not unsubstitutedand R₁ is not substituted by C₁₋₄alkyl.

[0066] More preferred R₁ groups wherein hydroxy is esterfied include:carboxypentanoyloxy, 4-carboxyphenylpropanoyloxy,4-(N-methylpiperizin-1-ylethyl)phenylcarbonyloxy,4-(piperizin-1-ylethyl)phenylcarbonyloxy,4-[N-di-(hydroxyethyl)aminomethyl]phenylcarbonyloxy,3-(N-acetylpiperizin-1-ylethyl)phenylcarbonyloxy,3-[N-di-(hydroxyethyl)aminomethyl]phenylcarbonyloxy,4-(N-methylpiperizin-1-ylpropanoylamino)phenylcarbonyloxy,N-methylpiperizin-1-ylcarbonylpropanoyloxy,N-di-(hydroxyethyl)aminocarbonylpropanoyloxy,piperizin-1-ylcarbonylpropanoyloxy,(N-acetylpiperizin-1-yl)carbonylpropanoyloxy,(N-di-(hydroxyethyl)aminocarbonylpropanoyloxy, and4-(piperizin-1-ylmethyl)phenylcarbonyloxy.

[0067] Further preferred R₁ groups wherein hydroxy is esterfied include:4-(N-methylpiperizin-1-ylpropanoylamino)phenylcarbonyloxy,N-methylpiperizin-1-ylcarbonylpropanoyloxy andN-di-(hydroxyethyl)aminocarbonylpropanoyloxy.

[0068] In this specification the generic term ‘alkyl’ includes bothstraight-chain and branched-chain alkyl groups. However references toindividual alkyl groups such as ‘propyl’ are specific for thestraight-chain version only and references to individual branched-chainalkyl groups such as ‘isopropyl’ are specific for the branched-chainversion only. An analogous convention applies to other generic terms.Examples of C₁₋₄alkyl include methyl, ethyl, propyl, isopropyl,sec-butyl and tert-butyl; Examples of C₁₋₄alkoxyC₁₋₄alkyl includepropxoymethyl, butoxyethyl or methoxyethyl; examples of aminoC₁₋₄alkylinclude aminomethyl, aminoethyl or aminopropyl; examples ofcarboxyC₁₋₄alkyl include carboxymethyl, carboxyethyl or carboxypropyl;example of carbamoylC₁₋₄alkyl include carbamoylethyl or carbamoylpropyl;examples of heterocyclylC₁₋₄alkyl include piperazinylmethyl,piperazinylethyl, morpholinylmethyl or morpholinylethyl; examples ofC₁₋₄alkoxy include methoxy, ethoxy and propoxy; examples ofN—C₁₋₄alkylamino include N-methylamino and N-ethylamino; examples ofN,N-di-C₁₋₄alkylamino include N,N-dimethylamino, N,N-diethylamino andN-methyl-N-ethylamino; examples of hydroxyC₁₋₄alkyl include hydroxyethylor hydroxypropyl; examples of C₁₋₄alkanoyl include formyl or propanoyl;examples of C₁₋₆alkanoyloxy include propanoyloxy or butanoyloxy;examples of C₁₋₄alkanoylC₁₋₄alkyl include formylethyl orpropanoylmethyl; examples of C₁₋₄alkanoylamino include acetylamino,propanoylamino and butanoylamino; examples ofheterocyclC₁₋₄alkanoylamino include piperizinylpropanoylamino orpiperidinylacetylamino; examples of C₁₋₄alkanoylheterocyclyl includeformylpiperidinyl, acetylpiperidinyl, formylpiperazinyl oracetylpiperazinyl; examples of arylcarbonyloxy includephenylcarbonyloxy; examples of heterocyclylcarbonyl includepiperazinylcarbonyl or piperidinylcarbonyl; examples ofheterocyclylcarbonyloxy include piperazinylcarbonyloxy,piperidinylcarbonyloxy or morpholinylcarbonyloxy; examples ofheteroarylcarbonyloxy include pyridinylcarbonyloxy orpyrimidinylcarbonyloxy; examples of C₁₋₄alkylthio includemethylsulfanyl, ethylsulfanyl and butylsulfanyl; examples ofphosphonoC₁₋₄alkyl include phosphonopropyl, phosphonoethyl orphosphonomethyl; and examples of carbamoylheterocyclyl includecarbamoylpiperazinyl or carbamoylpiperidinyl.

[0069] It is to be understood that, insofar as certain of the compoundsin the different features of the invention may exist in optically activeor racemic forms by virtue of one or more asymmetric carbon atoms, theinvention includes in its definition any such optically active orracemic form which possesses the property of inhibiting and/or reversingand/or alleviating the symptoms of angiogenesis and/or any diseasestates associated with angiogenesis. The synthesis of optically activeforms may be carried out by standard techniques of organic chemistrywell known in the art, for example by synthesis from optically activestarting materials or by resolution of a racemic form. Similarly,activity of these compounds may be evaluated using the standardlaboratory techniques referred to hereinafter.

[0070] The invention also relates to any and all tautomeric forms of thecompounds of the different features of the invention that possess theproperty of inhibiting and/or reversing and/or alleviating the symptomsof angiogenesis and/or any disease states associated with angiogenesis.

[0071] It will also be understood that certain compounds of the presentinvention may exist in solvated, for example hydrated, as well asunsolvated forms. It is to be understood that the present inventionencompasses all such solvated forms which possess the property ofinhibiting and/or reversing and/or alleviating the symptoms ofangiogenesis and/or any disease states associated with angiogenesis.

[0072] The compounds of Formula (I), Formula (II) or Formula (IIa) maybe administered in the form of a pro-drug which is broken down in thehuman or animal body to give a compound of the Formula (I), Formula (II)or Formula (IIa) respectively. Examples of pro-drugs include in-vivohydrolysable esters of a compound of the Formula (I), Formula (II) orFormula (IIa).

[0073] Various forms of pro-drugs are known in the art. For examples ofsuch pro-drug derivatives, see:

[0074] a) Design of Prodrugs, edited by H. Bundgaard, (Elsevier, 1985)and Methods in Enzymology, Vol. 42, p. 309-396, edited by K. Widder, etal. (Academic Press, 1985);

[0075] b) A Textbook of Drug Design and Development, edited byKrogsgaard-Larsen and H. Bundgaard, Chapter 5 “Design and Application ofProdrugs”, by H. Bundgaard p. 113-191 (1991);

[0076] c) H. Bundgaard, Advanced Drug Delivery Reviews, 8, 1-38 (1992);

[0077] d) H. Bundgaard, et al., Journal of Pharmaceutical Sciences, 77,285 (1988); and

[0078] e) N. Kakeya, et al., Chem Pharm Bull, 32, 692 (1984).

[0079] An in-vivo hydrolysable ester of a compound of the Formula (I),Formula (II) or Formula (IIa) containing a carboxy or a hydroxy groupis, for example, a pharmaceutically-acceptable ester which is hydrolysedin the human or animal body to produce the parent acid or alcohol.Suitable pharmaceutically-acceptable esters for carboxy includeC₁₋₆alkoxymethyl esters for example methoxymethyl, C₁₋₆alkanoyloxymethylesters for example pivaloyloxymethyl, phthalidyl esters,C₃₋₈cycloalkoxycarbonyloxyC₁₋₆alkyl esters for example1-cyclohexylcarbonyloxyethyl; 1,3-dioxolen-2-onylmethyl esters, forexample 5-methyl-1,3-dioxolen-2-onylmethyl; andC₁₋₆alkoxycarbonyloxyethyl esters.

[0080] An in-vivo hydrolysable ester of a compound of the Formula (I),Formula (II) or Formula (IIa) containing a hydroxy group includesinorganic esters such as phosphate esters (including phosphoramidiccyclic esters) and α-acyloxyalkyl ethers and related compounds which asa result of the in-vivo hydrolysis of the ester breakdown to give theparent hydroxy group/s. Examples of α-acyloxyalkyl ethers includeacetoxymethoxy and 2,2-dimethylpropionyloxy-methoxy. A selection ofin-vivo hydrolysable ester forming groups for hydroxy include alkanoyl,benzoyl, phenylacetyl and substituted benzoyl and phenylacetyl,alkoxycarbonyl (to give alkyl carbonate esters), dialkylcarbamoyl andN-(dialkylaminoethyl)-N-alkylcarbamoyl (to give carbamates),dialkylaminoacetyl and carboxyacetyl.

[0081] A suitable pharmaceutically-acceptable salt of a compound of theinvention is, for example, an acid-addition salt of a compound of theinvention which is sufficiently basic, for example, an acid-additionsalt with, for example, an inorganic or organic acid, for examplehydrochloric, hydrobromic, sulphuric, phosphoric, trifluoroacetic,citric or maleic acid. In addition a suitablepharmaceutically-acceptable salt of a benzoxazinone derivative of theinvention which is sufficiently acidic is an alkali metal salt, forexample a sodium or potassium salt, an alkaline earth metal salt, forexample a calcium or magnesium salt, an ammonium salt or a salt with anorganic base which affords a physiologically-acceptable cation, forexample a salt with methylamine, dimethylamine, trimethylamine,piperidine, morpholine or tris-(2-hydroxyethyl)amine.

[0082] A preferred group of values of X in each feature of the inventionis —O—, —S—, —S(O₂)—, or —N(R₄)—. Preferably X is —O—, —S— or —N(R₄)—.Most preferably X is —O— or —S—. In another embodiment of the inventionX is: —O—, —S—, —N(R₄)— or —N(R₄)CH₂C(O)—;

[0083] A preferred group of values of R₁ in each feature of theinvention is amino, hydroxy, methyl, C₁₋₄alkoxy or —OPO₃H₂ wherein theamino group is optionally substituted by an amino acid and the hydroxygroup is optionally esterified. Preferably R₁ is amino, hydroxy,methoxy, α-glutamylamino, serylamino, alanylamino or —OPO₃H_(2,) whereinthe hydroxy group is optionally esterified. More preferably R₁ is amino,hydroxy, methoxy, α-glutamylamino, serylamino, or —OPO₃H₂. In anotherembodiment of the invention R¹ is C₁₋₄alkyl, C₁₋₄alkoxy,C₁₋₄alkanoylamino, halo, amino, hydroxy, α-glutamylamino, or —OPO₃H₂.

[0084] A preferred group of values of R₂ in each feature of theinvention is hydrogen, methyl or ethyl; Preferably R₂ is hydrogen.

[0085] A preferred group of values of R₃ in each feature of theinvention is hydrogen, methyl or ethyl. Preferably R₃ is hydrogen.

[0086] A preferred group of values of R₄ in each feature of theinvention is hydrogen, methyl or ethyl. Preferably R₄ is hydrogen.

[0087] A preferred group of compounds of each feature of the invention,comprise compounds wherein:

[0088] X is —O—;

[0089] or a salt, pro-drug or solvate thereof.

[0090] A further preferred group of compounds of each feature of theinvention, comprise compounds wherein:

[0091] X is —N(R₄)—, preferably —NH—;

[0092] or a salt, pro-drug or solvate thereof.

[0093] A further preferred group of compounds of each feature of theinvention, comprise compounds wherein:

[0094] X is —S—, —S(O)— or —S(O₂)—, preferably —S—;

[0095] or a salt, pro-drug or solvate thereof.

[0096] A further preferred group of compounds of each feature of theinvention, comprise compounds wherein:

[0097] X is —N(R₄)CH₂C(O)—;

[0098] or a salt, pro-drug or solvate thereof.

[0099] A further preferred group of compounds of each feature of theinvention, comprise compounds wherein:

[0100] R₁ is amino, C₁₋₄alkoxy, hydroxy or —OPO₃H₂, wherein the aminogroup is optionally substituted by an amino acid residue and the hydroxygroup is optionally esterified;

[0101] or a salt, pro-drug or solvate thereof.

[0102] Particular compounds of each feature of the invention include:

[0103] 7-[(2-aminophenyl)sulfanyl]-2H-1,4-benzoxazin-3(4H)-one;

[0104] 7-(3-aminophenoxy)-2H-1,4-benzoxazin-3(4H)-one; and

[0105] 6-[2-(4-toluidino)acetyl]-2H-1,4-benzoxazin-3(4H)-one;

[0106] or a salt, pro-drug or solvate thereof.

[0107] More particular compounds of each feature of the inventioninclude:

[0108] 7-[(3-aminophenyl)sulfanyl]-2H-1,4-benzoxazin-3(4H)-one; and

[0109] 7-[(4-hydroxyphenyl)sulfanyl]-2H-1,4-benzoxazin-3(4H)-one;

[0110] or a salt, pro-drug or solvate thereof.

[0111] A compound of the invention or a pharmaceutically-acceptablesalt, pro-drug or solvate thereof, may be prepared by any process knownto be applicable to the preparation of chemically related compounds.Such processes, when used to prepare a compound of the invention or asalt, pro-drug or solvate thereof, are provided as a further feature ofthe invention and are illustrated by the following representativeexamples in which R₁, R₂, R₃, R₄, X, n and p have the same meaning asherein before defined. The reader is referred to Advanced OrganicChemistry, 4^(th) Edition, by Jerry March, published by John Wiley &Sons 1992, for general guidance on reaction conditions and reagents. Thereader is referred to Protective Groups in Organic Synthesis 2^(nd)Edition, by Green et al, published by John Wiley Sons for generalguidance on protecting groups.

[0112] Thus, according to the fourth feature of the invention there isprovided a process for preparing a compound of Formula (I), Formula (II)or Formula (IIa), or salt, pro-drug or solvate thereof, which process(wherein n, p, X, R₁, R₂, R₃, and R₄ are unless otherwise specified asdefined in Formula (I), Formula (II) or Formula (IIa) respectivelycomprises:

[0113] a) for compounds of Formula (I), Formula (I) or Formula (IIa)wherein X is —O—, —S— or —N(R₄)—, reacting a compound of Formula (A)with a compound of Formula (B),

[0114]  wherein L₁ is a leaving group;

[0115] b) for compounds of Formula (I), Formula (II) or Formula (IIa)wherein R₂ is hydrogen, reduction of a compound of Formula (C), whereinR₆ is hydrogen or an alkyl chain,

[0116] c) for compounds of Formula (I), Formula (II) or Formula (IIa)wherein R₂ is C₁₋₄alkyl, reacting a compound of Formula (I) wherein R₂is hydrogen with a suitable alkylhalide;

[0117] d) for compounds of Formula (I), Formula (II) or Formula (IIa)wherein X is —S(O)—, —S(O₂)—, oxidising a compound of Formula (D),

[0118] e) for compounds of Formula (I), Formula (II) or Formula (IIa)wherein X is —N(R₄)CH₂C(O)—, reacting a compound of Formula (E) with acompound of Formula (F),

[0119]  wherein L₂ is a leaving group;

[0120] and thereafter if necessary:

[0121] i) converting a compound of the Formula (I), Formula (II) orFormula (IIa) into another compound of the Formula (I), Formula (II) orFormula (IIa) respectively;

[0122] ii) removing any protecting groups;

[0123] iii) forming a salt, pro-drug or solvate.

[0124] According to a further aspect of the fourth feature of theinvention there is provided the processes a), b), c), d) and e)described above for the preparation of compounds of the Formula (I), ora salt, pro-drug or solvate thereof.

[0125] Specific reaction conditions for the above reactions are asfollows:

[0126] Process a) Compounds of Formula (A) and compound of Formula (B)can be reacted together in an organic solvent, at a temperature betweenroom temperature and about 80° C., optionally in the presence of a basesuch as sodium hydride, potassium carbonate or triethylamine.

[0127] Process b) The conditions for reduction of a compound of Formula(C) are well known in the art. Examples of reducing agents includehydrogen and a hydrogenation catalyst (for example palladium on carbon),zinc or iron in acetic acid or hydrochloric acid. The reaction ispreferably carried out in the presence of zinc with acetic acid, and ata temperature in the range of 0-80° C., preferably at or near roomtemperature.

[0128] Process c) Compounds of Formula (I), Formula (II) or Formula(IIa) wherein R₂ is hydrogen and a suitable alkylhalide may be reactedtogether in a suitable organic solvent such as DMF or DMSO, in thepresence of a base, such as sodium hydride or potassium carbonate at atemperature between about room temperature to about 80° C.

[0129] Process d) The oxidization of a compound of Formula (D) is wellknown in the art, for example, reaction with metachloroperbenzoic acid(MCPBA) in the presence of a suitable solvent such as dichloromethane atambient temperature. If an excess of MCPBA is used a compound of Formula(I), Formula (II) or Formula (IIa) wherein X is —S(O₂)— is obtained.

[0130] Process e) Compounds of Formula (E) and Formula (F) can bereacted together in the presence of an organic solvent, preferablyacetone, in the presence of a base, preferably potassium carbonate, at atemperature between about room temperature and 80° C.

[0131] Intermediates for the processes a), b), c) and d) can be preparedas outlined in Scheme 1, wherein P is a protecting group, using thefollowing reaction conditions:

[0132] Reaction Conditions (i) Reaction with ethyl bromoacetate in anorganic solvent such as DMF or acetone, in the presence of a base suchas sodium hydride or potassium carbonate at a temperature betweenapproximately room temperature and approximately 80° C.

[0133] Reaction Conditions (ii) Reduction using a suitable reducingagent such as hydrogen and a hydrogenation catalyst (for examplepalladium on carbon), iron or zinc in acetic acid or hydrochloric acid.

[0134] Reaction Conditions (iii) Reaction conditions for the removal ofa protecting group are well know in the art.

[0135] The compounds used as starting points for the reactions describedabove are commercially available or they are known compounds or they areprepared by processes known in the art.

[0136] It will also be appreciated that in some of the reactionsmentioned herein it may be necessary/desirable to protect any sensitivegroups in the compounds. The instances where protection is necessary ordesirable and suitable methods for protection are known to those skilledin the art. Conventional protecting groups may be used in accordancewith standard practice (for illustration see T. W. Green, ProtectiveGroups in Organic Synthesis, John Wiley and Sons, 1991). Thus, ifreactants include groups such as amino, carboxy or hydroxy it may bedesirable to protect the group in some of the reactions mentionedherein.

[0137] A suitable protecting group for an amino or alkylamino group is,for example, an acyl group, for example an alkanoyl group such asacetyl, an alkoxycarbonyl group, for example a methoxycarbonyl,ethoxycarbonyl or t-butoxycarbonyl group, an arylmethoxycarbonyl group,for example benzyloxycarbonyl, or an aroyl group, for example benzoyl.The deprotection conditions for the above protecting groups necessarilyvary with the choice of protecting group. Thus, for example, an acylgroup such as an alkanoyl or alkoxycarbonyl group or an aroyl group maybe removed for example, by hydrolysis with a suitable base such as analkali metal hydroxide, for example lithium or sodium hydroxide.Alternatively an acyl group such as a t-butoxycarbonyl group may beremoved, for example, by treatment with a suitable acid as hydrochloric,sulphuric or phosphoric acid or trifluoroacetic acid and anarylmethoxycarbonyl group such as a benzyloxycarbonyl group may beremoved, for example, by hydrogenation over a catalyst such aspalladium-on-carbon, or by treatment with a Lewis acid for example borontris(trifluoroacetate). A suitable alternative protecting group for aprimary amino group is, for example, a phthaloyl group which may beremoved by treatment with an alkylamine, for exampledimethylaminopropylamine, or with hydrazine.

[0138] A suitable protecting group for a hydroxy group is, for example,an acyl group, for example an alkanoyl group such as acetyl, an aroylgroup, for example benzoyl, or an arylmethyl group, for example benzyl.The deprotection conditions for the above protecting groups willnecessarily vary with the choice of protecting group. Thus, for example,an acyl group such as an alkanoyl or an aroyl group may be removed, forexample, by hydrolysis with a suitable base such as an alkali metalhydroxide, for example lithium or sodium hydroxide. Alternatively anarylmethyl group such as a benzyl group may be removed, for example, byhydrogenation over a catalyst such as palladium-on-carbon.

[0139] A suitable protecting group for a carboxy group is, for example,an esterifying group, for example a methyl or an ethyl group which maybe removed, for example, by hydrolysis with a base such as sodiumhydroxide, or for example a t-butyl group which may be removed, forexample, by treatment with an acid, for example an organic acid such astrifluoroacetic acid, or for example a benzyl group which may beremoved, for example, by hydrogenation over a catalyst such aspalladium-on-carbon.

[0140] The protecting groups may be removed at any convenient stage inthe synthesis using conventional techniques well known in the chemicalart.

[0141] In order to use a compound of the Formula (I), Formula (II) orFormula (IIa), or a pharmaceutically-acceptable salt, pro-drug orsolvate thereof, for the therapeutic treatment (including prophylactictreatment) of mammals including humans, it is normally formulated inaccordance with standard pharmaceutical practice as a pharmaceuticalcomposition.

[0142] The compositions of the invention may be in a form suitable fororal use (for example as tablets, lozenges, hard or soft capsules,aqueous or oily suspensions, emulsions, dispersible powders or granules,syrups or elixirs), for topical use (for example as creams, ointments,gels, or aqueous or oily solutions or suspensions), for administrationby inhalation (for example as a finely divided powder or a liquidaerosol), for administration by insufflation (for example as a finelydivided powder) or for parenteral administration (for example as asterile aqueous or oily solution for intravenous, subcutaneous,intramuscular or intramuscular dosing or as a suppository for rectaldosing).

[0143] The compositions of the invention may be obtained by conventionalprocedures using conventional pharmaceutical excipients, well known inthe art. Thus, compositions intended for oral use may contain, forexample, one or more colouring, sweetening, flavouring and/orpreservative agents.

[0144] Suitable pharmaceutically-acceptable excipients for a tabletformulation include, for example, inert diluents such as lactose, sodiumcarbonate, calcium phosphate or calcium carbonate, granulating anddisintegrating agents such as corn starch or algenic acid; bindingagents such as starch; lubricating agents such as magnesium stearate,stearic acid or talc; preservative agents such as ethyl or propylp-hydroxybenzoate, and anti-oxidants, such as ascorbic acid. Tabletformulations may be uncoated or coated either to modify theirdisintegration and the subsequent absorption of the active ingredientwithin the gastrointestinal tract, or to improve their stability and/orappearance, in either case, using conventional coating agents andprocedures well known in the art.

[0145] Compositions for oral use may be in the form of hard gelatincapsules in which the active ingredient is mixed with an inert soliddiluent, for example, calcium carbonate, calcium phosphate or kaolin, oras soft gelatin capsules in which the active ingredient is mixed withwater or an oil such as peanut oil, liquid paraffin, or olive oil.

[0146] Aqueous suspensions generally contain the active ingredient infinely powdered form together with one or more suspending agents, suchas sodium carboxymethylcellulose, methylcellulose,hydroxypropylmethylcellulose, sodium alginate, polyvinyl-pyrrolidone,gum tragacanth and gum acacia; dispersing or wetting agents such aslecithin or condensation products of an alkylene oxide with fatty acids(for example polyoxyethylene stearate), or condensation products ofethylene oxide with long chain aliphatic alcohols, for exampleheptadecaethyleneoxycetanol, or condensation products of ethylene oxidewith partial esters derived from fatty acids and a hexitol such aspolyoxyethylene sorbitol monooleate, or condensation products ofethylene oxide with partial esters derived from fatty acids and hexitolanhydrides, for example polyethylene sorbitan monooleate. The aqueoussuspensions may also contain one or more preservatives (such as ethyl orpropyl p-hydroxybenzoate, anti-oxidants (such as ascorbic acid),colouring agents, flavouring agents, and/or sweetening agents (such assucrose, saccharine or aspartame).

[0147] Oily suspensions may be formulated by suspending the activeingredient in a vegetable oil (such as arachis oil, olive oil, sesameoil or coconut oil) or in a mineral oil (such as liquid paraffin). Theoily suspensions may also contain a thickening agent such as beeswax,hard paraffin or cetyl alcohol. Sweetening agents such as those set outabove, and flavouring agents may be added to provide a palatable oralpreparation. These compositions may be preserved by the addition of ananti-oxidant such as ascorbic acid.

[0148] Dispersible powders and granules suitable for preparation of anaqueous suspension by the addition of water generally contain the activeingredient together with a dispersing or wetting agent, suspending agentand one or more preservatives. Suitable dispersing or wetting agents andsuspending agents are exemplified by those already mentioned above.Additional excipients such as sweetening, flavouring and colouringagents, may also be present.

[0149] The pharmaceutical compositions of the invention may also be inthe form of oil-in-water emulsions. The oily phase may be a vegetableoil, such as olive oil or arachis oil, or a mineral oil, such as forexample liquid paraffin or a mixture of any of these. Suitableemulsifying agents may be, for example, naturally-occurring gums such asgum acacia or gum tragacanth, naturally-occurring phosphatides such assoya bean, lecithin, an esters or partial esters derived from fattyacids and hexitol anhydrides (for example sorbitan monooleate) andcondensation products of the said partial esters with ethylene oxidesuch as polyoxyethylene sorbitan monooleate. The emulsions may alsocontain sweetening, flavouring and preservative agents.

[0150] Syrups and elixirs may be formulated with sweetening agents suchas glycerol, propylene glycol, sorbitol, aspartame or sucrose, and mayalso contain a demulcent, preservative, flavouring and/or colouringagent.

[0151] The pharmaceutical compositions may also be in the form of asterile injectable aqueous or oily suspension, which may be formulatedaccording to known procedures using one or more of the appropriatedispersing or wetting agents and suspending agents, which have beenmentioned above. A sterile injectable preparation may also be a sterileinjectable solution or suspension in a non-toxic parenterally-acceptablediluent or solvent, for example a solution in 1,3-butanediol.

[0152] Suppository formulations may be prepared by mixing the activeingredient with a suitable non-irritating excipient which is solid atordinary temperatures but liquid at the rectal temperature and willtherefore melt in the rectum to release the drug. Suitable excipientsinclude, for example, cocoa butter and polyethylene glycols.

[0153] Topical formulations, such as creams, ointments, gels and aqueousor oily solutions or suspensions, may generally be obtained byformulating an active ingredient with a conventional,topically-acceptable, vehicle or diluent using conventional procedureswell known in the art.

[0154] Compositions for administration by insufflation may be in theform of a finely divided powder containing particles of average diameterof, for example, 30 μm or much less, the powder itself comprising eitheractive ingredient alone or diluted with one or morephysiologically-acceptable carriers such as lactose. The powder forinsufflation is then conveniently retained in a capsule containing, forexample, 1 to 50 mg of active ingredient for use with a turbo-inhalerdevice, such as is used for insufflation of the known agent sodiumcromoglycate.

[0155] Compositions for administration by inhalation may be in the formof a conventional pressurised aerosol arranged to dispense the activeingredient either as an aerosol containing finely divided solid orliquid droplets. Conventional aerosol propellants such as volatilefluorinated hydrocarbons or hydrocarbons may be used and the aerosoldevice is conveniently arranged to dispense a metered quantity of activeingredient.

[0156] For further information on formulation the reader is referred toChapter 25.2 in Volume 5 of Comprehensive Medicinal Chemistry (CorwinHansch; Chairman of Editorial Board), Pergamon Press 1990.

[0157] The amount of active ingredient that is combined with one or moreexcipients to produce a single dosage form will necessarily varydepending upon the host treated and the particular route ofadministration. For example, a formulation intended for oraladministration to humans will generally contain, for example, from 0.5mg to 2 g of active agent compounded with an appropriate and convenientamount of excipients which may vary from about 5 to about 98 percent byweight of the total composition. Dosage unit forms will generallycontain about 1 mg to about 500 mg of an active ingredient. For furtherinformation on Routes of Administration and Dosage Regimes the reader isreferred to Chapter 25.3 in Volume 5 of Comprehensive MedicinalChemistry (Corwin Hansch; Chairman of Editorial Board), Pergamon Press1990.

[0158] The size of the dose for therapeutic or prophylactic purposes ofa compound of the Formula (I), Formula (II) or Formula (IIa) willnaturally vary according to the nature and severity of the conditions,the age and sex of the animal or patient and the route ofadministration, according to well known principles of medicine.

[0159] In using a compound of the Formula (I), Formula (II) or Formula(IIa) for therapeutic or prophylactic purposes it will generally beadministered so that a daily dose in the range of, for example, 0.5 mgto 75 mg per kg body weight is received, given if required in divideddoses. In general lower doses will be administered when a parenteralroute is employed. Thus, for example, for intravenous administration, adose in the range of, for example, 0.5 mg to 20 mg per kg body weightwill generally be used. Intranvenous administration is howeverpreferred, typically, intravenous doses of about 10 mg to 500 mg perpatient of a compound of this invention.

[0160] The compounds of this invention may be used in combination withother drugs and therapies used to inhibit and/or reverse and/oralleviate symptoms of angiogenesis and/or any disease state associatedwith angiogenesis. Examples of such disease states include: cancer,diabetes, psoriasis, rheumatoid arthritis, Kaposi's sarcoma,haemangioma, acute and chronic nephropathies, atheroma, arterialrestenosis, autoimmune diseases, acute inflammation, endometriosis,dysfunctional uterine bleeding and ocular diseases with retinal vesselproliferation.

[0161] If formulated as a fixed dose such combination products employthe compounds of this invention within the dosage range described hereinand the other pharmaceutically-active agent within its approved dosagerange. Sequential use is contemplated when a combination formulation isinappropriate.

[0162] In the field of medical oncology examples of such combinationsinclude combinations with the following categories of therapeutic agent:

[0163] i) anti-angiogenic agents that work by different mechanisms fromthe compounds of Formula (I), Formula (II) or Formula (IIa) (for examplelinomide, inhibitors of integrin αvβ3 function, angiostatin, endostatin,razoxin, thalidomide) and including vascular endothelial growth factor(VEGF) receptor tyrosine kinase inhibitors (RTKIs) (for example thosedescribed in international patent applications publication nos.WO-97/22596, WO-97/30035, WO-97/32856 and WO-98/13354, the entiredisclosure of which documents is incorporated herein by reference);

[0164] ii) cytostatic agents such as anti-oestrogens (for exampletamoxifen, toremifene, raloxifene, droloxifene, iodoxyfene),progestogens (for example megestrol acetate), aromatase inhibitors (forexample anastrozole, letrazole, vorazole, exemestane),anti-progestogens, anti-androgens (for example flutamide, nilutamide,bicalutamide, cyproterone acetate), LHRH agonists and antagonists (forexample goserelin acetate, luprolide), inhibitors of testosterone5α-dihydroreductase (for example finasteride), anti-invasion agents (forexample metalloproteinase inhibitors like marimastat and inhibitors ofurokinase plasminogen activator receptor function) and inhibitors ofgrowth factor function, (such growth factors include for exampleepidermal growth factor (EGF), platelet derived growth factor andhepatocyte growth factor such inhibitors include growth factorantibodies, growth factor receptor antibodies, tyrosine kinaseinhibitors and serine/threonine kinase inhibitors);

[0165] iii) biological response modifiers (for example interferon);

[0166] iv) antibodies (for example edrecolomab); and

[0167] v) anti-proliferative/anti-neoplastic drugs and combinationsthereof, as used in medical oncology, such as anti-metabolites (forexample anti-folates like methotrexate, fluoropyrimidines like5-fluorouracil, purine and adenosine analogues, cytosine arabinoside);anti-tumour antibiotics (for example anthracyclines like doxorubicin,daunomycin, epirubicin and idarubicin, mitomycin-C, dactinomycin,mithramycin); platinum derivatives (for example cisplatin, carboplatin);alkylating agents (for example nitrogen mustard, melphalan,chlorambucil, busulphan, cyclophosphamide, ifosfamide, nitrosoureas,thiotepa); anti-mitotic agents (for example vinca alkaloids likevincristine and taxoids like taxol, taxotere); enzymes (for exampleasparaginase); thymidylate synthase inhibitors (for exampleraltitrexed); topoisomerase inhibitors (for example epipodophyllotoxinslike etoposide and teniposide, amsacrine, topotecan, irinotecan).

[0168] The compounds of the invention may also be used in combinationwith surgery or radiotherapy.

[0169] According to the fifth feature of the present invention there isprovided a compound of Formula (I), Formula (II) or Formula (IIa), orpharmaceutically-acceptable salt, pro-drug or solvate thereof,preferably in the form of a pharmaceutical composition, which when dosedin divided doses (also known as split doses) produces a greateranti-tumour effect than when a single dose is given.

[0170] Anti-tumour effects include but are not limited to, inhibition oftumour growth, tumour growth delay, regression of tumour, shrinkage oftumour, increased time to re-growth of tumour on cessation of treatment,slowing of disease progression. It is expected that when a compound ofthe present invention is administered to a warm-blooded animal such as ahuman, in need of treatment for cancer involving a solid tumour, saidmethod of treatment will produce an effect, as measured by, for example,one or more of: the extent of the anti-tumour effect, the response rate,the time to disease progression and the survival rate.

[0171] According to a further aspect of the fifth feature of the presentinvention there is provided a method for the production of a vasculardamaging effect in a warm-blooded animal such as a human, whichcomprises administering to said animal in divided doses an effectiveamount of a compound of Formula (I), Formula (II) or Formula (IIa), orpharmaceutically-acceptable salt, pro-drug or solvate thereof,preferably in the form of a pharmaceutical composition.

[0172] According to a further aspect of the fifth feature of the presentinvention there is provided a method for the treatment of a cancerinvolving a solid tumour in a warm-blooded animal such as a human, whichcomprises administering to said animal in divided doses an effectiveamount of a compound of Formula (I), Formula (II) or Formula (IIa), orpharmaceutically-acceptable salt, pro-drug or solvate thereof,preferably in the form of a pharmaceutical composition.

[0173] According to a further aspect of the fifth feature of the presentinvention there is provided a medicament comprising two or morefractions of doses of a compound of Formula (I), Formula (II) or Formula(IIa), or pharmaceutically-acceptable salt, pro-drug or solvate thereof,preferably in the form of a pharmaceutical composition, which togetheradd up to a total daily dose, for administration in divided doses foruse in a method of treatment of a human or animal body by therapy.

[0174] According to a further aspect of the fifth feature of the presentinvention there is provided a kit comprising two or more fractions ofdoses of a compound of Formula (I), Formula (II) or Formula (IIa), orpharmaceutically-acceptable salt, pro-drug or solvate thereof,preferably in the form of a pharmaceutical composition, which togetheradd up to a total daily dose, for administration in divided doses.

[0175] According to a further aspect of the fifth feature of the presentinvention there is provided a kit comprising:

[0176] a) two or more fractions of doses of a compound of Formula (I),Formula (II) or Formula (IIa), or pharmaceutically-acceptable salt,pro-drug or solvate thereof, which together add up to a total dailydose, in unit dosage forms for administration in divided doses; and

[0177] b) container means for containing said dosage forms.

[0178] According to a further aspect of the fifth feature of the presentinvention there is provided a kit comprising:

[0179] a) two or more fractions of doses of a compound of Formula (I),Formula (II) or Formula (IIa), or pharmaceutically-acceptable salt,pro-drug or solvate thereof, which together add up to a total dailydose, together with an excipient or carrier, in unit dosage forms; and

[0180] b) container means for containing said dosage forms.

[0181] According to a further aspect of the fifth feature of the presentinvention there is provided the use of a compound of Formula (I),Formula (II) or Formula (IIa), or pharmaceutically-acceptable salt,pro-drug or solvate thereof, in the manufacture of a medicament foradministration in divided doses for use in the production of a vasculardamaging effect in a warm-blooded animal such as a human.

[0182] According to a further aspect of the fifth feature of the presentinvention there is provided the use of a compound of Formula (I),Formula (II) or Formula (IIa), or pharmaceutically-acceptable salt,pro-drug or solvate thereof, in the manufacture of a medicament foradministration in divided doses for use in the production of ananti-cancer effect in a warm-blooded animal such as a human.

[0183] According to a further aspect of the fifth feature of the presentinvention there is provided the use of a compound of Formula (I),Formula (II) or Formula (IIa), or pharmaceutically-acceptable salt,pro-drug or solvate thereof, in the manufacture of a medicament foradministration in divided doses for use in the production of ananti-tumour effect in a warm-blooded animal such as a human.

[0184] Divided doses, also called split doses, means that the total doseto be administered to a warm-blooded animal, such as a human, in any oneday period (for example one 24 hour period from midnight to midnight) isdivided up into two or more fractions of the total dose and thesefractions are administered with a time period between each fraction ofabout greater than 0 hours to about 10 hours, preferably about 1 hour toabout 6 hours, more preferably about 2 hours to about 4 hours. Thefractions of total dose may be about equal or unequal.

[0185] Preferably the total dose is divided into two parts which may beabout equal or unequal.

[0186] The time intervals between doses may be for example selectedfrom: about 1 hour, about 1.5 hours, about 2 hours, about 2.5 hours,about 3 hours, about 3.5 hours, about 4 hours, about 4.5 hours, about 5hours, about 5.5 hours and about 6 hours.

[0187] The time intervals between doses may be any number (includingnon-integers) of minutes between greater than 0 minutes and 600 minutes,preferably between 45 and 375 minutes inclusive. If more than two dosesare administered the time intervals between each dose may be about equalor unequal.

[0188] Preferably two doses are given with a time interval in betweenthem of greater than or equal to 1 hour and less than 6 hours.

[0189] More preferably two doses are given with a time interval inbetween them of greater than or equal to two hours and less than 5hours.

[0190] Yet more preferably two doses are given with a time interval inbetween them of greater than or equal to two hours and less than orequal to 4 hours.

[0191] Particularly the total dose is divided into two parts which maybe about equal or unequal with a time interval between doses of greaterthan or equal to about two hours and less than or equal to about 4hours.

[0192] More particularly the total dose is divided into two parts whichmay be about equal with a time interval between doses of greater than orequal to about two hours and less than or equal to about 4 hours.

[0193] For the avoidance of doubt the term ‘about’ in the description oftime periods means the time given plus or minus 15 minutes, thus forexample about 1 hour means 45 to 75 minutes, about 1.5 hours means 75 to105 minutes. Elsewhere the term ‘about’ has its usual dictionarymeaning.

[0194] Although the compounds of the Formula (I), Formula (II) orFormula (IIa) are primarily of value as therapeutic agents for use inwarm-blooded animals (including man), they are also useful whenever itis required to inhibit and/or reverse and/or alleviate symptoms ofangiogenesis and/or any disease state associated with angiogenesis.Thus, they are useful as pharmacological tools for use in thedevelopment of new biological tests and in the search for newpharmacological agents.

[0195] Biological Assay

[0196] Colchicine Binding Site Competitive Assay Kit

[0197] The ability of a ligand to bind specifically to the colchicinebinding site on tubulin, an indicator of the vascular damaging activity,was assessed using a size exclusion chromatography assay kit from“Cytoskeleton” (1650 Fillmore St. #240, Denver, Colo. 80206, U.S.A.)Catalogue number of kit: BK023.

[0198] The following reagents were used:

[0199] tubulin buffer, to give 0.1 mM GTP, 0.5 mM MgCl₂, 0.5 mM EGTA, 40mM PIPES buffer at pH 6.9 in the final reaction mix;

[0200] purified tubulin protein from bovine brain at 1 mg/ml in tubulinbuffer;

[0201] 0.02 mM fluorescent colchicine in tubulin buffer [FITC(fluorescein isothiocyanate)-labelled];

[0202] 2 mM colchicine in tubulin buffer;

[0203] 0.2 mM vinblastine in tubulin buffer; and

[0204] G-25 Sephadex™ Fine—particle size 34-138 μm.

[0205] The reaction was performed as follows:

[0206] 8 μl of test compound (dissolved in DMSO) was gently mixed with150 μl of tubulin. This was then incubated at 37° C. for 30 minutes.Then 4 μl of the fluorescent colchicine was added, the incubation mixvortexed for 5 seconds and then incubated for a further 30 minutes at37° C. At the end of the reaction incubation size exclusionchromatography was performed to separate the tubulin with fluorescentcolchicine bound from the free, unbound colchicine. If a test compoundinhibited fluorescent colchicine binding then a reduced signal ismeasured and the compound is confirmed as a colchicine site bindingmoiety.

[0207] Size exclusion chromatography was performed as follows, usingchromatography columns filled with 3 mls of G-25 Sephadex™ Fine slurry.The incubation mixture was pipetted onto the column and up to 12elutions of 160 μl were collected. The fluorescence of thetubulin-containing fractions was detected on a spectrophotometer whichexcites at 485 nm and emits at 535 nm.

[0208] Control incubations were also performed, 8 μl DMSO (negativecontrol) and 8 μl colchicine stock (positive competition control),instead of the 8 μl of test compound in the incubation mixture.

[0209] The degree of competition of colchicine binding by eitherunlabelled colchicine or test compound was calculated relative to theDMSO negative control.

[0210] Compounds of Formula (I), Formula (II) or Formula (IIa) encompassvascular damaging agents and pro-drugs of vascular damaging agents.Pro-drugs of vascular damaging agents are believed to be cleavedin-vivo. Without being bound by theoretical considerations thesepro-drugs may have lower activity in the in-vitro colchicine bindingsite competitive assay, than would be anticipated when the activity ofthese compounds is measured in cell based assays or in-vivo.

[0211] The invention will now be illustrated with the followingnon-limiting Examples in which, unless otherwise stated:

[0212] (i) evaporations were carried out by rotary evaporation in vacuoand work-up procedures were carried out after removal of residual solidssuch as drying agents by filtration;

[0213] (ii) operations were carried out at ambient temperature, that isin the range 18-25° C. and under an atmosphere of an inert gas such asargon or nitrogen;

[0214] (iii) yields are given for illustration only and are notnecessarily the maximum attainable;

[0215] (iv) the structures of the end-products of the Formula (I),Formula (II) or Formula (IIa) were confirmed by nuclear (generallyproton) magnetic resonance (NMR) and mass spectral techniques; protonmagnetic resonance chemical shift values were measured on the deltascale and peak multiplicities are shown as follows: s, singlet; d,doublet; t, triplet; m, multiplet; br, broad; q, quartet, quin, quintet;

[0216] (v) intermediates were not generally fully characterised andpurity was assessed by thin layer chromatography (TLC), high-performanceliquid chromatography (HPLC), infra-red (IR) or NMR analysis;

[0217] (vi) flash chromatography was performed on silica (MerckKeiselgel: Art.9385);,

[0218] (vii) OASIS™ is a macroporous co-polymer, used to purifyhydrophilic compounds, made from a balanced ratio of lipophilicdivinylbenzene and hydrophilic N-vinylpyrrolidone. OASIS™ is describedin the following patents, U.S. Pat. No. 5,882,521, U.S. patent No.5,976,376 and U.S. Pat. No. 6,106,721. OASIS™ sample extraction productswere obtained from Waters Corporation (Milford, Mass., USA).Abbreviations 4-Dimethylaminopyridine DMAP1-(3-Dimethylanminopropyl)-3-ethylcarbodiimide EDCI hydrochlorideDimethyl sulphoxide DMSO N-(9-fluorenylmethoxycarbonyl) N-FMOC

EXAMPLE 1 7-(4-Methoxyphenoxy)-2H-1,4-benzoxazin-3(4H)-one

[0219]

[0220] A solution of 1 (1.2 g; 3.4 mmol) in AcOH (22 ml) and water (3ml) was treated portionwise with zinc (2.3 g; 34 mmol). The mixture wasstirred at room temperature for 1 hour. After evaporation to dryness theresidue was taken up in EtOAc, filtered and the solid washed well withEtOAc. The filtrate was washed with water, then brine and dried overMgSO₄. The residue was purified by flash chromatography eluting withincreasingly polar mixtures of EtOAc/hexanes (40 to 50% EtOAc) to give7-(4-methoxyphenoxy)-2H-1,4-benzoxazin-3(4H)-one as a beige solid (0.71g).

[0221] Yield: 76% M. Pt. 163-164° C. ¹H NMR spectrum (DMSO d₆): 3.74 (s,3H); 4.55 (s, 2H); 6.54 (m, 2H); 6.85 (d, 1H); 6.95 (m, 4H); 10.65 (brs, 1H). LCMS-ESI: 272 [MH]⁺ Elemental analysis: Found C 66.66 H 4.8 N5.24 C₁₅H₁₁N₃OS Requires C 66.41 H 4.83 N 5.16

[0222] The starting material was prepared as follows:—

[0223] A solution of 2 (31.4 g; 200 mmol) in acetone (300 ml) wastreated with K₂CO₃ (33 g; 240 mmol) and warmed to 40° C. A solution ofethyl bromoacetate (35 g; 210 mmol) in acetone (30 ml) was added over 5minutes and the mixture was stirred and heated at 70° C. for 2 hour 30minutes and allowed to cool. The mixture was evaporated to dryness,taken up in Et₂O and the organic phase washed with water, brine anddried over MgSO₄. Evaporation and trituration with hexanes gave 3 as apale yellow solid (47.0 g).

[0224] Yield: 97% ¹H NMR spectrum (CDCl₃): 1.30 (t, 3H); 4.29 (q, 2H);4.77 (s, 2H); 6.69 (dd, 1H); 6.81 (ddd, 1H); 7.98 (dd, 1H). LCMS-ESI:244 [M]⁺

[0225] Compound 4 (1.24 g; 10 mmol) was dissolved inN,N-dimethylacetamide (10 ml) under argon and was treated with NaH (450mg; 60% in oil; 11 mmol). The mixture was stirred for 1 hour and asolution of compound 3 (2.43 g; 10 mmol) was added. The mixture washeated at 60° C. and stirred for 20 hours and then allowed to cool. Themixture was poured into water and extracted with EtOAc (3×). The organicphase was washed with water, saturated. aqueous. NaHCO₃, then brine anddried over MgSO₄.

[0226] The residue (3 g) was purified by flash chromatography elutingwith increasingly polar mixtures of EtOAc/hexanes (20 to 30% EtOAc). Theappropriate fractions were evaporated to give 1 as a yellow oil (2.9 g).

[0227] Yield: 84% ¹H NMR spectrum (CDCl₃): 1.28 (t, 3H); 3.83 (s, 3H);4.24 (q, 2H); 4.69 (s, 2H); 6.46 (d, 1H); 6.51 (dd, 1H); 6.97 (m, 4H);7.94 (d, 1H). LCMS-ESI: 348 [MH]⁺

EXAMPLE 2 7-(4-Hydroxyphenoxy)-2H-1,4-benzoxazin-3(4H)-one

[0228]

[0229] A solution of 7-(4-methoxyphenoxy)-2H-1,4-benzoxazin-3(4H)-one(120 mg; 0.44 mmol) in CH₂Cl₂ (1 ml) was treated with BBr₃ (332 mg; 1.33mmol). The mixture was stirred at room temperature for 3 hours. Afterevaporation to dryness the residue was taken up in EtOAc and water. Theorganic phase was washed with brine and dried over MgSO₄. The residue(110 mg) was triturated with EtOAc, filtered and dried to give7-(4-hydroxyphenoxy)-2H-1,4-benzoxazin-3(4H)-one as a beige solid (85mg), Melting point 250-251° C.

[0230] Yield: 75% ¹H NMR spectrum (DMSO d₆): 4.54(s, 2H); 6.51 (m, 2H);6.75 (d, 2H); 6.84 (m, 3H); 9.31 (s, 1H); 10.62 (s, 1H). LCMS-ESI: 258[MH]⁺ Elemental analysis: Found C 64.74 H 4.51 N 5.29 C₁₄H₁₁NO₄; 0.1 H₂ORequires C 64.91 H 4.36 N 5.41

EXAMPLE 3 7-[(4-Aminophenyl)sulfanyl]-2H-1,4-benzoxazin-3(4H)-one

[0231]

[0232] Using the general method described for Example 1,7-[(4-aminophenyl)sulfanyl]-2H-1,4-benzoxazin-3(4H)-one was obtainedfrom 5 (2.0 g; 5.7 mmol) after purification by flash chromatographyeluting with increasingly polar mixtures of EtOAc/hexanes (70 to 100%EtOAc). The product was a beige solid (1.41 g), Melting point 232-234°C.

[0233] Yield: 90% ¹H NMR spectrum (DMSO d₆): 4.51 (s, 2H); 5.48 (s, 2H);6.58 (m, 3H); 6.70 (dd, 1H); 6.79 (d, 1H); 7.14 (m, 2H); 10.66 (s, 1H).LCMS-ESI: 271 [M−H]⁻ Elemental analysis: Found C 61.90 H 4.44 N 10.29 S11.29 C₁₄H₁₂N₂O₂S Requires C 61.75 H 4.44 N 10.29 S 11.77

[0234] The starting material was prepared as follows:—

[0235] Compound 3 (2.43 g; 10 mmol) and compound 6 (1.25 g; 10 mmol)were dissolved in N-methyl-2-pyrrolidinone (20 ml) under argon and weretreated with K₂CO₃ (2.1 g; 15 mmol). The mixture was heated at 80° C.and stirred for 2.5 hours and then allowed to cool. The mixture waspoured into water and extracted with EtOAc (3×). The organic phase waswashed with water, brine and dried over MgSO₄.

[0236] The residue (3.6 g) was purified by flash chromatography elutingwith increasingly polar mixtures of EtOAc/hexanes (40 to 50% EtOAc). Theappropriate fractions were evaporated to give 5as a pale beige solid(2.1 g), Melting point 110-111° C.

[0237] Yield: 60% ¹H NMR spectrum (CDCl₃): 1.28 (t, 3H); 3.97 (s, 2H);4.22 (q, 2H); 4.62 (s, 2H); 6.53 (d, 1H); 6.67 (dd, 1H); 6.72 (m, 2H);7.30 (m, 2H); 7.78 (d, 1H). LCMS-ESI: 349 [MH]⁺ Elemental analysis:Found C 55.5 H 4.67 N 8.16 S 8.41 C₁₆H₁₆N₂O₅S Requires C 55.16 H 4.63 N8.04 S 9.2

EXAMPLE 4 7-[(2-Aminophenyl)sulfanyl]-2H-1,4-benzoxazin-3(4H)-one

[0238]

[0239] Using the general method described for Example 1,7-[(2-aminophenyl)sulfanyl]-2H-1,4-benzoxazin-3(4H)-one was obtainedfrom 7 (3.5 g; 10 mmol) after purification by flash chromatographyeluting with increasingly polar mixtures of EtOAc/hexanes (40 to 50%EtOAc). The product was a white solid (1.05 g), Melting point 204-205°C.

[0240] Yield: 39% ¹H NMR spectrum (DMSO d₆): 4.54 (s, 2H); 5.38 (s, 2H);6.59 (t, 1); 6.64 (d, 1H); 6.8 (m, 3H); 7.17 (t, 1H); 7.3 (dd, 1H); 10.7(s, 1H). LCMS-ESI: 271 [M−H]⁻ Elemental analysis: Found C 62.20 H 4.51 N10.32 S 11.35 C₁₄H₁₂N₂O₂S Requires C 61.75 H 4.44 N 10.29 S 11.77

[0241] The starting material was prepared as follows:—

[0242] Using the general method described for the starting material inExample 3, except that the mixture was heated for 3 hours, 7 wasobtained from 3 (2.43 g; 10 mmol) and 8 (1.25 g; 10 mmol) afterpurification by flash chromatography eluting with increasingly polarmixtures of EtOAc/hexanes (20 to 25% EtOAc). The appropriate fractionswere evaporated to give 7 as a pale yellow solid (3.7 g), Melting point70-71° C.

[0243] Yield: 100% ¹H NMR spectrum (CDCl₃): 1.27 (t, 3H); 4.20 (q, 2H);4.28 (s, 2H); 4.61 (s, 2H); 6.51 (d, 1H); 6.71 (dd, 1H); 6.79 (dt, 1H);6.83 (dd, 1H); 7.31 (dt, 1H); 7.41 (dd, 1H); 7.81 (d, 1H). LCMS-ESI: 349[MH]⁺ Elemental analysis: Found C 55.42 H 4.76 N 8.15 S 8.36 C₁₆H₁₆N₂O₅SRequires C 55.16 H 4.63 N 8.04 S 9.2

EXAMPLE 5 7-[(3-Aminophenyl)sulfanyl]-2H-1,4-benzoxazin-3(4H)-one

[0244]

[0245] Using the general method described for Example 1,7-[(3-aminophenyl)sulfanyl]-2H-1,4-benzoxazin-3(4H)-one was obtainedfrom 9 (8.65 g; 25 mmol) after purification by flash chromatographyeluting with increasingly polar mixtures of EtOAc/hexanes (50 to 70%EtOAc). The product was a beige solid (3.05 g), Melting point 181-182°C.

[0246] Yield: 45% ¹H NMR spectrum (DMSO d₆): 4.59 (s, 2H); 5.22 (s, 2H);6.41 (m, 1H); 6.5 (m, 2H); 6.9 (m, 2H); 7.0 (m, 2H); 10.82 (s, 1H).LCMS-ESI: 271 [M−H]⁻ Elemental analysis: Found C 61.87 H 4.51 N 10.13 S11.52 C₁₄H₁₂N₂O₂S Requires C 61.75 H 4.44 N 10.29 S 11.77

[0247] The starting material was prepared as follows:—

[0248] Using the general method described for the starting material inExample 3, except that the mixture was heated for 3 hours, 9 wasobtained from 3 (9.72 g; 40 mmol) and 10 (5.0 g; 40 mmol) afterpurification by flash chromatography eluting with increasingly polarmixtures of EtOAc/hexanes (30 to 50% EtOAc). The appropriate fractionswere evaporated to give 9 as a yellow oil (13.78 g).

[0249] Yield: 99% ¹H NMR spectrum (CDCl₃): 1.27 (t, 3H); 3.81 (s, 2H);4.22 (q, 2H); 4.64 (s, 2H); 6.67 (d, 1H); 6.72 (m, 1H); 6.79 (m, 2H);6.86 (m, 1H); 7.20 (t, 1H); 7.8 (d, 1H). LCMS-ESI: 349 [MH]⁺ Elementalanalysis: Found C 55.36 H 4.78 N 7.83 S 8.76 C₁₆H₁₆N₂O₅S Requires C55.16 H 4.63 N 8.04 S 9.2

EXAMPLE 6 7-[(4-Hydroxyphenyl)sulfanyl]-2H-1,4-benzoxazin-3(4H)-one

[0250]

[0251] Using the general method described for Example 1,7-[(4-Hydroxyphenyl)sulfanyl]-2H-1,4-benzoxazin-3(4H)-one was obtainedfrom 11 (1.9 g; 5.4 mmol) after purification by flash chromatographyeluting with increasingly polar mixtures of EtOAc/hexanes (50 to 70%EtOAc). The product was a beige solid (1.32 g), Melting point 224-225°C.

[0252] Yield: 89% ¹H NMR spectrum (DMSO d₆): 4.54 (s, 2H); 6.67 (d, 1H);6.8 (m, 4H); 7.27 (d, 2H); 9.82 (br s, 1H); 10.71 (s, 1H). LCMS-ESI: 272[M−H]⁻ Elemental analysis: Found C 61.53 H 4.17 N 5.18 S 11.03C₁₄H₁₁NO₃S Requires C 61.53 H 4.06 N 5.12 S 11.73

[0253] The starting material was prepared as follows:—

[0254] Using the general method described for the starting material inExample 3, 11 was obtained from 3 (2.43 g; 10 mmol) and 12 (1.26 g; 10mmol) after purification by flash chromatography eluting withincreasingly polar mixtures of EtOAc/hexanes (30 to 50% EtOAc). Theappropriate fractions were evaporated to give 11 as a pale yellow solid(2.48 g), Melting point 156-157° C.

[0255] Yield: 71% ¹H NMR spectrum (CDCl₃): 1.28 (t, 3H); 4.21 (q, 2H);4.61 (s, 2H); 6.50 (d, 1H); 6.68 (dd, 1H); 6.94 (d, 2H); 7.36 (d, 2H);7.78 (d, 1H); 9.28 (s, 1H). LCMS-ESI: 348 [M−H]⁻ Elemental analysis:Found C 55.12 H 4.45 N 4.07 S 8.82 C₁₆H₁₅NO₆S Requires C 55.01 H 4.33 N4.01 S 9.18

EXAMPLE 7 7-[(3,4-Dimethoxyphenyl)sulfanyl]-2H-1,4-benzoxazin-3(4H)-one

[0256]

[0257] Using the general method described for Example 1,7-[(3,4-dimethoxyphenyl)sulfanyl]-2H-1,4-benzoxazin-3(4H)-one wasobtained from 13 (3.34 g; 8.5 mmol) after purification by flashchromatography eluting with increasingly polar mixtures of EtOAc/hexanes(50 to 70% EtOAc). The product was a pink solid (1.95 g). A smallquantity was recrystallised from EtOAc, Melting point 170-171° C.

[0258] Yield: 73% ¹H NMR spectrum (DMSO d₆): 3.75 (s, 3H); 3.78 (s, 3H);4.57 (s, 2H); 6.78 (d, 1H); 6.87 (m, 2H); 7.0 (m, 3H); 10.76 (s, 1H).LCMS-ESI: 316 [M−H]⁻ Elemental analysis: Found C 60.18 H 4.94 N 4.34 S9.23 C₁₆H₁₅NO₄S Requires C 60.55 H 4.76 N 4.41 S 10.10

[0259] The starting material was prepared as follows:—

[0260] Using the general method described for the starting material inExample 3, 13 was obtained from 3 (2.43 g; 10 mmol) and 14 (1.7 g; 10mmol) after purification by flash chromatography eluting withincreasingly polar mixtures of EtOAc/hexanes (30 to 50% EtOAc). Theappropriate fractions were evaporated to give 13 as a yellow solid (3.34g), Melting point 97-98° C.

[0261] Yield: 85% ¹H NMR spectrum (CDCl₃): 1.27 (t, 3H); 3.88 (s, 3H);3.94 (s, 3H); 4.20 (q, 2H); 4.62 (s, 2H); 6.55 (d, 1H); 6.70 (dd, 1H);6.94 (d, 1H); 7.0 (d, 1H); 7.15 (dd, 1H); 7.80 (d, 1H). LCMS-ESI: 394[MH]⁺ Elemental analysis: Found C 55.17 H 4.92 N 3.58 S 7.09 C₁₈H₁₉NO₇SRequires C 54.95 H 4.87 N 3.56 S 8.15

EXAMPLE 8 7-[(3,4-Dihydroxyphenyl)sulfanyl]-2H-1,4-benzoxazin-3(4H)-one

[0262]

[0263] A solution of Example 7(7-[(3,4-dimethoxyphenyl)sulfanyl]-2H-1,4-benzoxazin-3(4H)-one)(320 mg;1.0 mmol) in CH₂Cl₂ (2 ml) was treated with BBr₃ (750 mg; 3.0 mmol). Themixture was stirred at room temperature for 3 hours. After evaporationto dryness the residue was taken up in EtOAc and water. The organicphase was washed with water, brine and dried over MgSO4. The residue(110 mg) was triturated with EtOAc/CH₂Cl₂, filtered and dried to give7-[(3,4-dihydroxyphenyl)sulfanyl]-2H-1,4-benzoxazin-3(4H)-one as a beigesolid (280 mg), Melting point 236-238° C.

[0264] Yield: 96% ¹H NMR spectrum (DMSO d₆): 4.54(s, 2H); 6.69 (d, 1H);6.74-6.84 (m, 5H); 9.22 (s, 1H); 9.27 (s, 1H); 10.72 (s, 1H). LCMS-ESI:288 [M−H]⁻ Elemental analysis: Found C 57.98 H 4.10 N 4.63 S 10.77C₁₄H₁₁NO₄S Requires C 58.12 H 3.83 N 4.84 S 11.08

EXAMPLE 9 7-Benzylsulfanyl-2H-1,4-benzoxazin-3(4H)-one

[0265]

[0266] Using the general method described for Example 1,7-benzylsulfanyl-2H-1,4-benzoxazin-3(4H)-one was obtained from 15 (1.5g; 4.3 mmol) after purification by flash chromatography eluting withincreasingly polar mixtures of EtOAc/hexanes (30 to 50% EtOAc). Theproduct was a beige solid (1.11 g), Melting point 154-155° C.

[0267] Yield: 95% ¹H NMR spectrum (DMSO d₆): 4.15 (s, 2H); 4.55 (s, 2H);6.80 (d, 1H); 6.92 (m, 2H); 7.21-7.32 (m, 5H); 10.71 (s, 1H). LCMS-ESI:270 [M−H]⁻ Elemental analysis: Found C 66.15 H 4.86 N 5.27 S 10.34C₁₅H₁₃NO₂S Requires C 66.40 H 4.83 N 5.16 S 11.82

[0268] The starting material was prepared as follows:—

[0269] Compounds 3 (1.215 g; 5 mmol) and 16 (0.62 g; 5 mmol) weredissolved in DMF (5 ml) and treated with triethylamine (0.85 ml; 6mmol). The mixture was stirred and heated at 70° C. for 3 days and thenallowed to cool. The mixture was poured into EtOAc and the organic phasewashed with aqueous NaOH (1N), water then brine and dried over MgSO₄.Evaporation gave essentially pure 15 (1.69 g). A small quantity waspurified by flash chromatography eluting with increasingly polarmixtures of EtOAc/hexanes (20 to 40% EtOAc). The appropriate fractionswere evaporated to give a yellow solid, Melting point 76-77° C.

[0270] Yield: 99% ¹H NMR spectrum (CDCl₃): 1.29 (t, 3H); 4.20 (s, 2H);4.26 (q, 2H); 4.65 (s, 2H); 6.77 (d, 1H); 6.93 (dd, 1H); 7.35 (m, 5H);7.83 (d, 1H). LCMS-ESI: 348 [MH]⁺ Elemental analysis: Found C 58.06 H4.89 N 4.15 S 7.22 C₁₇H₁₇NO₅S, 0.2 H₂O Requires C 58.17 H 5.00 N 3.99 S9.14

EXAMPLE 10 7-(4-Chlorobenzylsulfanyl)-2H-1,4-benzoxazin-3(4H)-one

[0271]

[0272] Using the general method described for Example 1,7-(4-chloro-benzylsulfanyl)-2H-1,4-benzoxazin-3(4H)-one was obtainedfrom 17 (1.7 g; 4.4 mmol) after purification by flash chromatographyeluting with increasingly polar mixtures of EtOAc/hexanes (30 to 50%EtOAc). The product was a cream solid (1.04 g), Melting point 174-175°C.

[0273] Yield: 76% ¹H NMR spectrum (DMSO d₆): 4.15 (s, 2H); 4.55 (s, 2H);6.79 (d, 1H); 6.90 (dd, 1H); 6.93 (d, 1H); 7.32 (m, 4H); 10.72 (s, 1H).LCMS-ESI: 304.5 [M−H]⁻ Elemental analysis: Found C 58.93 H 4.05 N 4.67 S9.90 C₁₅H₁₂ClNO₂S Requires C 58.92 H 3.96 N 4.58 S 10.49

[0274] The starting material was prepared as follows:—

[0275] Using the general method described for the starting material inExample 9, 17 (1.89 g) was obtained from 3 (1.215 g; 5 mmol) and 18(0.79 g; 5 mmol). A small quantity was purified by flash chromatographyeluting with increasingly polar mixtures of EtOAc/hexanes (30 to 50%EtOAc). The appropriate fractions were evaporated to give 17 as a yellowsolid, Melting point 97-98° C.

[0276] Yield: 99% ¹H NMR spectrum (CDCl₃): 1.30 (t, 3H); 4.16 (s, 2H);4.27 (q, 2H); 4.68 (s, 2H); 6.78 (d, 1H); 6.93 (dd, 1H); 7.31 (m, 4H);7.85 (d, 1H). LCMS-ESI: 382.5 [MH]⁺ Elemental analysis: Found C 53.25 H4.29 N 3.77 S 7.29 C₁₇H₁₆ClNO₅S Requires C 53.48 H 4.22 N 3.67 S 8.40

EXAMPLE 11 7-(4-tert-Butylbenzylsulfanyl)-2H-1,4-benzoxazin-3(4H)-one

[0277]

[0278] Using the general method described for Example 1,7-(4-tert-butyl-benzylsulfanyl)-2H-1,4-benzoxazin-3(4H)-one was obtainedfrom 19 (1.8 g; 4.4 mmol) after purification by flash chromatographyeluting with increasingly polar mixtures of EtOAc/hexanes (30 to 50%EtOAc). The product was a beige solid (1.23 g), Melting point 134-136°C.

[0279] Yield: 84% ¹H NMR spectrum (DMSO d₆): 1.25 (s, 9H); 4.13 (s, 2H);4.55 (s, 2H); 6.80 (d, 1H); 6.93 (m, 2H); 7.29 (m, 4H); 10.71 (s, 1H).LCMS-ESI: 326 [M−H]⁻ Elemental analysis: Found C 69.55 H 6.57 N 4.31 S8.79 C₁₉H₂₁NO₂S Requires C 69.69 H 6.46 N 4.28 S 9.79

[0280] The starting material was prepared as follows:—

[0281] Using the general method described for the starting material inExample 9, 19 (2.0 g) was obtained from 3 (1.215 g; 5 mmol) and 18 (0.9g; 5 mmol). A small quantity was purified by flash chromatographyeluting with increasingly polar mixtures of EtOAc/hexanes (30 to 50%EtOAc). The appropriate fractions were evaporated to give 19 as a yellowsolid, Melting point 98-99° C.

[0282] Yield: 99% ¹H NMR spectrum (CDCl₃): 1.27 (t, 3H); 1.31 (s, 9H);4.17 (s, 2H); 4.26 (q, 2H); 4.64 (s, 2H); 6.77 (d, 1H); 6.94 (dd, 1H);7.33 (m, 4H); 7.84 (d, 1H). LCMS-ESI: 404 [MH]⁺ Elemental analysis:Found C 62.65 H 6.36 N 3.75 S 6.71 C₂₁H₂₅NO₅S Requires C 62.51 H 6.25 N3.47 S 7.95

EXAMPLE 12 7-(4-Methoxybenzylsulfanyl)-2H-1,4-benzoxazin-3(4H)-one

[0283]

[0284] Using the general method described for Example 1,7-(4-methoxy-benzylsulfanyl)-2H-1,4-benzoxazin-3(4H)-one was obtainedfrom 21 (1.89 g ; 5.0 mmol) after trituration with EtOAc. The productwas a beige solid (1.32 g).

[0285] Yield: 87% ¹H NMR spectrum (DMSO d₆): 3.71 (s, 3H); 4.10 (s, 2H);4.55 (s, 2H); 6.80 (d, 1H); 6.84 (m, 2H); 6.91 (dd, 1H); 6.92 (d, 1H);7.22 (m, 2H); 10.70 (s, 1H). LCMS-ESI: 300 [M−H]* Elemental analysis:Found C 63.58 H 5.03 N 4.72 S 10.27 C₁₆H₁₅NO₃S Requires C 63.77 H 5.02 N4.65 S 10.64

[0286] The starting material was prepared as follows:—

[0287] Using the general method described for the starting material inExample 9, 21 (1.89 g) was obtained from 3 (1.215 g; 5 mmol) and 22(0.77 g; 5 mmol).

[0288] Yield: 99% ¹H NMR spectrum (CDCl₃): 1.29 (t, 3H); 3.80 (s, 3H);4.15 (s, 2H); 4.26 (q, 2H); 4.67 (s, 2H); 6.77 (d, 1H); 6.86 (m, 2H);6.93 (dd, 1H); 7.27 (m, 2H); 7.83 (d, 1H). LCMS-ESI: 378 [MH]⁺

EXAMPLE 13 7-(4-Hydroxybenzylsulfanyl)-2H-1,4-benzoxazin-3(4H)-one

[0289]

[0290] A solution of7-(4-methoxy-benzylsulfanyl)-2H-1,4-benzoxazin-3(4H)-one (150 mg; 0.5mmol) in CH₂Cl₂ (2 ml) was treated with BBr₃ (350 mg; 1.5 mmol). Themixture was stirred at room temperature overnight. After evaporation todryness the residue was taken up in EtOAc and water. The organic phasewas washed with water, brine and dried over MgSO4. The residue wastriturated with EtOAc, filtered and dried to give7-(4-hydroxy-benzylsulfanyl)-2H-1,4-benzoxazin-3(4H)-one as a beigesolid (149 mg), Melting point 207-209° C.

[0291] Yield: 100% ¹H NMR spectrum (DMSO d₆): 4.07(s, 2H); 4.56(s, 2H);6.68 (m, 2H); 6.81 (d, 1H); 6.91 (m, 2H); 7.11 (d, 2H); 9.35 (s, 1H);10.72 (s, 1H). LCMS-ESI: 288 [MH]⁺ Elemental analysis: Found C 62.24 H4.77 N 4.81 S 10.01 C₁₅H₁₃NO₃S; Requires C 62.23 H 4.83 N 4.59 S 10.510.2 EtOAc

EXAMPLE 14 7-(4-Methoxyanilino)-2H-1,4-benzoxazin-3(4H)-one

[0292]

[0293] Using the general method described for Example 1,7-(4-methoxyanilino)-2H-1,4-benzoxazin-3(4H)-one was obtained from 23(500 mg; 1.4 mmol) after purification by flash chromatography elutingwith increasingly polar mixtures of EtOAc/hexanes (50 to 60% EtOAc). Theproduct was a beige solid (363 mg), Melting point 174-175° C.

[0294] Yield: 93% ¹H NMR spectrum (DMSO d₆): 3.72 (s, 3H); 4.50 (s, 2H);6.52 (d, 1H); 6.55 (dd, 1H); 6.74 (d, 1H); 6.86 (m, 2H); 6.99 (m, 2H);7.73 (s, 1H); 10.46 (s, 1H). LCMS-ESI: 271 [MH]⁺ Elemental analysis:Found C 66.50 H 5.14 N 10.31 C₁₅H₁₄N₂O₃ Requires C 66.66 H 5.22 N 10.36

[0295] The starting material was prepared as follows:—

[0296] Compound 3 (1.21 g; 5 mmol) and compound 24 (0.61 g; 5 mmol) weredissolved in N,N-dimethylacetamide (10 ml) under argon and the mixturewas heated at 90° C. and stirred for 14 hours. The mixture was furtherheated at 100° C. for 3 days and then allowed to cool. The mixture waspoured into water and extracted with EtOAc (3×). The organic phase waswashed with water, brine and dried over MgSO₄.

[0297] The residue (1.75 g) was purified by flash chromatography elutingwith increasingly polar mixtures of EtOAc/hexanes (20 to 40% EtOAc). Theappropriate fractions were evaporated to give 23 as a gum (500 mg).

[0298] Yield: 30% ¹H NMR spectrum (CDCl₃): 1.26 (t, 3H); 3.83 (s, 3H);4.23 (q, 2H); 4.65 (s, 2H); 6.15 (s, 1H); 6.24 (d, 1H); 6.39 (dd, 1H);6.92 (m, 2H); 7.12 (m, 2H); 7.96 (d, 1H). LCMS-ESI: 347 [MH]⁺

EXAMPLE 15 7-(4-Hydroxyanilino)-2H-1,4-benzoxazin-3(4H)-one

[0299]

[0300] A solution of 7-(4-methoxyanilino)-2H-1,4-benzoxazin-3(4H)-one(135 mg; 0.5 mmol) in CH₂Cl₂ (2 ml) was treated with BBr₃ (350 mg; 1.5mmol). The mixture was stirred at room temperature for overnight. Afterevaporation to dryness the residue was taken up in EtOAc and saturatedaqueous NaHCO₃. The organic phase was washed with water, brine and driedover MgSO₄. The residue (147 mg) was triturated with EtOAc, filtered anddried to give 7-(4-hydroxyanilino)-2H-1,4-benzoxazin-3(4H)-one as amagenta solid (120 mg), Melting point 270-275° C.

[0301] Yield: 94% ¹H NMR spectrum (DMSO d₆): 4.48(s, 2H); 6.46 (d, 1H);6.48 (dd, 1H); 6.68-6.72 (m, 3H); 6.89 (m, 2H); 7.56 (s, 1H); 8.98 (s,1H); 10.42 (s, 1H). LCMS-ESI: 256 [MH]⁺ Elemental analysis: Found C65.07 H 4.83 N 10.55 C₁₄H₁₂N₂O₃; 0.1 EtOAc Requires C 65.25 H 4.78 N10.57

EXAMPLE 16 7-(4-Aminophenoxy)-2H-1,4-benzoxazin-3(4H)-one

[0302]

[0303] Using the general method described for Example 1,7-(4-aminophenoxy)-2H- 1,4-benzoxazin-3(4H)-one was obtained from 25(690 mg; 2.0 mmol) after purification by flash chromatography elutingwith increasingly polar mixtures of EtOAc/hexanes (70 to 100% EtOAc).The product was a white solid (243 mg), Melting point 201-202° C.

[0304] Yield: 46% ¹H NMR spectrum (DMSO d₆): 4.52 (s, 2H); 4.95 (s, 2H);6.44 (d, 1H); 6.45 (dd, 1H); 6.56 (m, 2H); 6.73 (m, 2H); 6.80 (d, 1H);10.58 (s, 1H). LCMS-ESI: 255 [M−H]⁻ Elemental analysis: Found C 65.06 H4.58 N 10.85 C₁₄H₁₂N₂O₃; 0.1 H₂O Requires C 65.16 H 4.77 N 10.86

[0305] The starting material was prepared as follows:—

[0306] Compound 26 (1.09 g; 10 mmol) was dissolved inN,N-dimethylacetamide (10 ml) under argon and was treated with NaH (450mg; 60% in oil; 11 mmol). The mixture was stirred for 1 hour and asolution of compound 3 (2.43 g; 10 mmol) was added. The mixture washeated at 60° C. and stirred for 2 hours then further heated at 85° C.for 3.5 hours and then allowed to cool. The mixture was poured intowater and extracted with EtOAc (3×). The organic phase was washed withwater, saturated aqueous NaHCO₃, then brine and dried over MgSO₄. Theresidue (3 g) was purified by flash chromatography eluting withincreasingly polar mixtures of EtOAc/hexanes (50 to 60% EtOAc). Theappropriate fractions were evaporated to give 25 as a yellow oil (690mg).

[0307] Yield: 20% ¹H NMR spectrum (CDCl₃): 1.28 (t, 3H); 3.70 (s, 2H);4.25 (q, 2H); 4.69 (s, 2H); 6.46 (d, 1H); 6.51 (dd, 1H); 6.71 (m, 2H);6.87 (m, 2H); 7.94 (d, 1H). LCMS-ESI: 333 [MH]⁺

EXAMPLE 17 7-(3-Aminophenoxy)-2H-1,4-benzoxazin-3(4H)-one

[0308]

[0309] Using the general method described for Example 1,7-(3-aminophenoxy)-2H-1,4-benzoxazin-3(4H)-one was obtained from 27 (800mg; 2.4 mmol) after purification by flash chromatography eluting withincreasingly polar mixtures of EtOAc/hexanes (50 to 70% EtOAc). Theproduct was a white solid (239 mg), Melting point 209-210° C.

[0310] Yield: 39% ¹H NMR spectrum (DMSO d₆): 4.56 (s, 2H); 5.19 (s, 2H);6.09 (m, 2H); 6.14 (t, 1H); 6.29 (m, 1H); 6.59 (m, 2H); 6.86 (m, 1H);6.96 (t, 1H); 10.66 (s, 1H). LCMS-ESI: 255 [M−H]⁻ Elemental analysis:Found C 65.99 H 4.72 N 11.02 C₁₄H₁₂N₂O₃ Requires C 65.62 H 4.72 N 10.93

[0311] The starting material was prepared as follows:—

[0312] Using the general method described for the starting material inExample P, 27 (1.0 g) was obtained as a yellow oil from 3 (2.43 g; 10mmol) and 28 (1.09 g; 10 mmol) after purification by flashchromatography eluting with increasingly polar mixtures of EtOAc/hexanes(40 to 50% EtOAc).

[0313] Yield: 30% ¹H NMR spectrum (CDCl₃): 1.28 (t, 3H); 3.82 (s, 2H);4.25 (q, 2H); 4.70 (s, 2H); 6.36 (t, 1H); 6.41 (dd, 1H); 6.5-6.6 (m,3H); 7.16 (t, 1H); 7.94 (d, 1H). LCMS-ESI: 333 [MH]⁺

EXAMPLE 18 7-(2-Aminophenoxy)-2H-1,4-benzoxazin-3(4H)-one

[0314]

[0315] Using the general method described for Example 1,7-(2-aminophenoxy)-2H-1,4-benzoxazin-3(4H)-one was obtained from 29 (400mg; 1.2 mmol) after purification by flash chromatography eluting withincreasingly polar mixtures of EtOAc/hexanes (50 to 70% EtOAc). Theproduct was a light beige solid (201 mg), Melting point 149-150° C.

[0316] Yield: 60% ¹H NMR spectrum (DMSO d₆): 4.53 (s, 2H); 4.87 (s, 2H);6.49-6.55 (m, 3H); 6.75-6.85 (m, 3H); 6.90 (t, 1H); 10.61 (s, 1H).LCMS-ESI: 255 [M−H]⁻ Elemental analysis: Found C 64.93 H 4.86 N 10.78C₁₄H₁₂N₂O₃; 0.2 H₂O Requires C 64.71 H 4.81 N 10.78

[0317] The starting material was prepared as follows:—

[0318] Using the general method described for the starting material inExample 16, 29 (1.93 g) was obtained as a yellow oil from 3 (2.43 g; 10mmol) and 30 (1.09 g; 10 mmol) after purification by flashchromatography eluting with increasingly polar mixtures of EtOAc/hexanes(20 to 40% EtOAc).

[0319] Yield: 60% ¹H NMR spectrum (CDCl₃): 1.27 (t, 3H); 3.74 (s, 2H);4.23 (q, 2H); 4.69 (s, 2H); 6.50 (d, 1H); 6.57 (dd, 1H); 6.77 (dt, 1H);6.85 (dd, 1H); 6.92 (dd, 1H); 7.08 (dt, 1H); 7.95 (d, 1H). LCMS-ESI: 333[MH]⁺

EXAMPLE 19 7-(4-Acetylaminophenoxy)-2H-1,4-benzoxazin-3(4H)-one

[0320]

[0321] Using the general method described for Example 1,7-(4-acetylaminophenoxy)-2H-1,4-benzoxazin-3(4H)-one was obtained from31 (980 mg; 2.6 mmol) after purification by flash chromatography elutingwith increasingly polar mixtures of EtOAc/hexanes (80 to 100% EtOAc).The product was a pale cream solid (770 mg), Melting point 230-232° C.

[0322] Yield: 99% ¹H NMR spectrum (DMSO d₆): 2.02; (s, 3H); 4.56 (s,2H); 6.57 (m, 2H); 6.86 (dd, 1H); 6.93 (m, 2H); 7.56 (m, 2H); 9.92 (s,1H); 10.66 (s, 1H). LCMS-ESI: 297 [M−H]⁻ Elemental analysis: Found C63.46 H 5.01 N 9.16 C₁₆H₁₄N₂O₄; 0.2 MeOH Requires C 63.86 H 4.90 N 9.19

[0323] The starting material was prepared as follows:—

[0324] Using the general method described for the starting material inExample 3, except that N,N-dimethylacetamide was used as solvent and themixture was heated at 110° C. for 18 hours, 31 (1.85 g) was obtained asa pale yellow oil from 3 (7.29 g; 30 mmol) and 32 (4.5 g; 30 mmol) afterpurification by flash chromatography eluting with increasingly polarmixtures of EtOAc/hexanes (60 to 80% EtOAc).

[0325] Yield: 16% ¹H NMR spectrum (CDCl₃): 1.27 (t, 3H); 2.20 (s, 3H);4.25 (q, 2H); 4.70 (s, 2H); 6.53 (m, 2H); 7.03 (d, 2H); 7.31 (s, 1H);7.56 (d, 2H); 7.95 (d, 1H). LCMS-ESI: 375 [MH]⁺ Elemental analysis:Found C 56.92 H 4.82 N 7.37 C₁₈H₁₈N₂O₇; 0.3 H₂O Requires C 56.93 H 4.94N 7.38

EXAMPLE 20 6-[2-(4-Toluidino)acetyl]-2H-1,4-benzoxazin-3(4H)-one

[0326]

[0327] A solution of 33 (226 mg; 1.0 mmol) and 34(118 mg; 1.1 mmol) inacetone (10 ml) was treated with K₂CO₃ (276 mg; 2.0 mmol) and then KI(166 mg; 1.0 mmol). The mixture was stirred at room temperature for 2days. Water was added and the precipitate filtered, washed with waterand dried. The residue was was purified by flash chromatography elutingwith CH₂Cl₂/EtOAc/MeOH (70/30/5). The appropriate fractions wereevaporated to give 6-[2-(4-toluidino)acetyl]-2H-1,4-benzoxazin-3(4H)-oneas a beige solid (60 mg).

[0328] Yield: 20% ¹H NMR spectrum (DMSO d₆): 2.16(s, 3H); 4.56(d, 2H);4.71(s, 2H); 5.60 (t, 1H); 6.58 (d, 2H); 6.91 (d, 2H); 7.09 (d, 1H);7.54 (d, 1H); 7.78 (dd, 1H); 10.88 (s, 1H). LCMS-ESI: 297 [MH]⁺

EXAMPLE 21 6-[2-(4-Aminoanilino)acetyl]-2H-1,4-benzoxazin-3(4H)-one

[0329]

[0330] Compound 35 (200 mg; 0.5 mmol) was taken up in a saturatedsolution of gaseous HCl in EtOAc (10 ml) and the mixture stirred for 3days. The precipitate that remained was filtered washed with Et₂O anddried to give 6-[2-(4-aminoanilino)acetyl]-2H-1,4benzoxazin-3(4H)-one asa beige solid (180 mg).

[0331] Yield: 100% ¹H NMR spectrum (DMSO d₆): 4.62 (s, 2H); 4.70 (s,2H); 6.72 (d, 2H); 7.09 (m, 3H); 7.53 (d, 1H); 7.76 (dd, 1H); 9.75 (brs, 3H); 10.90 (s, 1H). LCMS-ESI: 296 [M−H]⁻ Elemental analysis: Found C51.25 H 4.75 N 11.05 Cl 16.5 C₁₆H₁₅N₃O₃, Requires C 51.42 H 4.90 N 11.24Cl 16.60 1.75 HCl, 0.7 H₂O

[0332] The starting material was prepared as follows:—

[0333] Compound 36 (1.04 g; 5 mmol) was dissolved in acetone (40 ml) andwas treated with K₂CO₃ (1.38 g; 10 mmol) and KI (830 mg; 5 mmol). Themixture was stirred for 1 hour. A suspension of 33 (1.41 g; 5.25 mmol)in acetone (20 ml) was added and the mixture was heated under reflux for1 hour. The cooled mixture was poured into water and extracted withEtOAc (3×). The organic phase was washed with water, brine and driedover MgSO₄. The residue was purified by trituration in hot MeOH to give35 as a yellow solid (1.16 g).

[0334] Yield: 58% ¹H NMR spectrum (CDCl₃): 1.44 (s, 9H); 4.54 (d, 2H);4.69 (s, 2H); 5.53 (t, 1H); 6.58 (m, 2H); 7.08 (d, 1H); 7.14 (m, 2H);7.53 (d, 1H); 7.75 (dd, 1H); 8.82 (s, 1H); 10.87 (s, 1H). LCMS-ESI: 398[MH]⁺

EXAMPLE 22 7-(4-L-a-Glutamyl-phenylsulfanyl)-2H-1,4-benzoxazin-3(4H)-one

[0335]

[0336] A solution of 37 (160 mg; 0.35 mmol) in EtOAc (3 ml) was treatedwith a saturated solution of gaseous HCl in EtOAc (10 ml). The mixturewas stirred at room temperature overnight. After evaporation to dryness,the residue was taken up in water and purified by reverse phasechromatography on an OASIS resin eluting with increasingly lipophilicmixtures of MeCN/water (0 to 100% MeCN), buffered with 1% aqueous HCl.The appropriate fractions were evaporated and dried to give thehydrochloride of7-(4-L-a-glutamyl-benzylsulfanyl)-2H-1,4-benzoxazin-3(4H)-one as a whitesolid (113 mg).

[0337] Yield: 72% ¹H NMR spectrum (DMSO d₆, CD₃OD): 2.09 (m, 2H); 2.42(m, 2H); 4.04 (m, 1H); 4.56 (s, 2H); 6.86 (d, 1H); 6.92 (m, 2H); 7.32(d, 2H); 7.63 (d, 2H); 10.78 (s, 1M). LCMS-ESI: 400 [M−H]⁻

[0338] The starting material was prepared as follows:—

[0339] A solution of Example 3(7-[(4-aminophenyl)sulfanyl]-2H-1,4-benzoxazin-3(4H)-one) (275 mg; 1.0mmol) and 38 (850 mg; 2.0 mmol) in CH₂Cl₂ (10 ml) was treated with EDCI(480 mg; 2.5 mmol) and a catalytic quantity of DMAP (10 mg). The mixturewas stirred at room temperature for 1 hour. The mixture was directlypurified by flash chromatography eluting with CH₂Cl₂/EtOAc (50/50). Theappropriate fractions were evaporated to give 39 as a white solid (648mg).

[0340] Yield: 95% ¹H NMR spectrum (CDCl₃): 1.46 (s, 9H); 2.03 (m, 1H);2.17 (m, 1H); 2.41 (m, 1H); 2.57 (m, 1H); 4.20 (t, 1H); 4.40 (m, 3H);4.58 (s, 2H); 6.08 (m, 1H); 6.72 (d, 1H); 6.90 (m,2H); 7.26 (m, 2H);7.31 (d, 2H); 7.38 (t, 2H); 7.51 (d, 2H); 7.57 (t, 2H); 7.74 (d, 2H);8.67 (s, 1H); 8.84 (s, 1H).

[0341] A solution of 39 (1.08 g; 1.4 mmol) in CH₂Cl₂ (40 ml) and DMF (5ml) was treated with piperidine (20 ml). The mixture was stirred at roomtemperature for 1 hour. After evaporation to dryness, the residue waspurified by flash chromatography eluting with increasingly polarmixtures of MeOH/CH₂Cl₂ (0 to 10% MeOH). The appropriate fractions wereevaporated to give 37 as a white solid (389 mg).

[0342] Yield: 53% ¹H NMR spectrum (DMSO d₆): 1.40 (s, 9H); 1.67 (m, 1H);1.86 (m, 1H); 2.32 (m, 2H); 3.32 (m, 1H); 4.57 (s, 2H); 6.82 (d, 1H);6.90 (m, 2H); 7.32 (d, 2H); 7.68 (d, 2H); 10.78 (s, 1H). LCMS-ESI: 456[M−H]⁻

EXAMPLE 23 7-(3-L-a-Glutamyl-phenylsulfanyl)-2H-1,4-benzoxazin-3(4H)-one

[0343]

[0344] Using the general method described in Example 22,7-(3-L-a-glutamyl-phenylsulfanyl)-2H-1,4-benzoxazin-3(4H)-one (563 mg)was obtained as a white solid from 40 (640 mg; 1.4 mmol) afterpurification by reverse phase chromatography eluting with increasinglylipophilic mixtures of MeCN/water (0 to 100% MeCN), buffered with 1%aqueous HCl.

[0345] Yield: 99% ¹H NMR spectrum (DMSO d₆): 2.07 (m, 2H); 2.39 (m, 2H);4.01 (m, 1H); 4.61 (s, 2H); 6.97 (m, 3H); 7.07 (dd, 1H); 7.33 (t, 1H);7.50 (d, 1H); 7.58 (s, 1H); 8.37 (m, 2H); 10.81 (s, 1H); 10.91 (s, 1H).LCMS-ESI: 400 [M−H]⁻ Elemental analysis: Found C 49.99 H 4.79 N 9.29 S6.66 C₁₉H₁₉N₃O₅S; Requires C 50.24 H 4.70 N 9.25 S 7.06 1.2 HCl 0.5 H₂O

[0346] The starting material was prepared as follows:—

[0347] Using the general method described for the starting material inExample 22, 42 (2.43 g) was obtained as a white solid from Example 5(7-[(3-Aminophenyl)sulfanyl]-2H-1,4-benzoxazin-3(4H)-one (820 mg; 3.0mmol) and 41 (2.3 g; 5.4 mmol) after purification by flashchromatography eluting with increasingly polar mixtures of EtOAc/hexanes(40 to 70% EtOAc).

[0348] Yield: 100% ¹H NMR spectrum (CDCl₃): 1.45 (s, 9H); 2.01 (m, 1H);2.13 (m, 1H); 2.38 (m, 1H); 2.51 (m, 1H); 4.17 (t, 1H); 4.38 (m, 3H);4.55 (s, 2H); 6.00 (m, 1H); 6.71 (d, 1H); 6.97 (dd, 1H); 7.00 (d, 1H);7.05 (d, 1H); 7.24 (m, 3H); 7.37 (t, 2H); 7.48 (d, 2H); 7.55 (t, 2H);7.73 (d, 2H); 8.60 (s, 1H); 8.76 (s, 1H). LCMS-ESI: 680 [MH]⁺

[0349] and similarly 40 (1.17 g) was obtained as a white solid from 42(2.4 g; 3.5 mmol) after purification by flash chromatography elutingwith increasingly polar mixtures of MeOH/CH₂Cl₂ (0 to 10% MeOH).

[0350] Yield: 73% ¹H NMR spectrum (DMSO d₆): 1.40 (s, 9H); 1.65 (m, 1H);1.85 (m, 1H); 2.29 (m, 2H); 3.30 (m, 1H); 4.61 (s, 2H); 6.96 (m, 3H);7.03 (dd, 1H); 7.29 (t, 1H); 7.53 (d, 2H) 7.65 (s, 1H); 10.85 (s, 1H).LCMS-ESI: 456 [M−H]⁻

EXAMPLE 24 7-(4-Phosphonophenoxy)-2H-1,4-benzoxazin-3(4H)-one

[0351]

[0352] A solution of 43 (310 mg; 0.6 mmol) in EtOAc (5 ml) and MeOH (5ml) was hydrogenated at 30 psi at room temperature for 30 minutes. Afterfiltration and evaporation to dryness, the residue was taken up in waterand purified by reverse phase chromatography on an OASIS resin elutingwith increasingly lipophilic mixtures of MeCN/water (0 to 50% MeCN),buffered with 1% aqueous HCl. The appropriate fractions were adjusted topH 7.0 with aqueous NaOH (0.1N) and freeze-dried to give the disodiumsalt of 7-(4-phosphonophenoxy)-2H-1,4-benzoxazin-3(4H)-one as a whitesolid (200 mg).

[0353] Yield: 87% ¹H NMR spectrum (DMSO d₆, CD₃OD): 4.52 (s, 2H); 6.56(m, 2H); 6.88 (m, 3H); 7.14 (d, 2H); 10.54 (s, 1H). LCMS-ESI: 336 [M−H]⁻Elemental analysis: Found C 42.30 H 3.11 N 3.63 C₁₄H₁₀NO₇P; 1.8 Na 1.2H₂O Requires C 42.19 H 3.15 N 3.51

[0354] The starting material was prepared as follows:—

[0355] Example 2 (7-(4-hydroxyphenoxy)-2H-1,4-benzoxazin-3(4H)-one) (514mg; 2 mmol) was dissolved in CH₃CN (20 ml) under argon and was treatedsuccessively with CCl₄ (1 ml), diisopropylethylamine (730 ul; 4.2 mmol),DMAP (10 mg) and dibenzyl phosphite (1.320 ml; 6 mmol). The mixture wasstirred at room temperature for 24 hours. The mixture was evaporated,poured into aqueous. KH₂PO₄ and extracted with CH₂Cl₂.

[0356] The residue was purified by flash chromatography eluting withincreasingly polar mixtures of EtOAc/CH₂Cl₂ (20 to 30% EtOAc). Theappropriate fractions were evaporated to give 43 as a white solid (570mg).

[0357] Yield: 55% ¹H NMR spectrum (DMSO d₆): 4.60 (s, 2H); 5.16 (s, 2H);5.18 (s, 2H); 6.64 (m, 2H); 6.91 (d, 1H); 7.00 (m, 2H); 7.20 (m, 2H);7.39 (m, 10H); 10.72 (s, 1H). LCMS-ESI: 516 [M−H]⁻

1. A compound of Formula (II), wherein:

X is selected from: —O—, —S—, —S—(O)—, —S(O₂)—, —N(R₄)— or—N(R₄)CH₂C(O)—; R₁ is independently selected from: amino, halo, hydroxy,—OPO₃H₂, C₁₋₄alkyl, C₁₋₄alkoxy, N—C₁₋₄alkylamino, N,N-di-C₁₋₄alkylamino,C_(1,4)alkanoylamino or C₁₋₄alkylthio wherein the amino group isoptionally substituted by an amino acid residue and the hydroxy group isoptionally esterified; R₂ is selected from: hydrogen or C₁₋₄alkyl; R₃ isselected from: hydrogen or C₁₋₄alkyl; R₄ is selected from: hydrogen orC₁₋₄alkyl; n is 0, 1 or 2; and p is 0, 1, 2 or 3; with the proviso thatthe following compounds are excluded:6-benzyloxy-2H-1,4-benzoxazin-3(4H)-one;7-benzyloxy-2H-1,4-benzoxazin-3(4H)-one;2-methyl-7-benzyloxy-2H-1,4-benzoxazin-3(4H)-one;2,4-dimethyl-7-benzyloxy-2H-1,4-benzoxazin-3(4H)-one;2-methyl-7-(3,5-dichlorobenzyloxy)-2H-1,4-benzoxazin-3(4H)-one;2,4-dimethyl-7-(3,5-dichlorobenzyloxy)-2H-1,4-benzoxazin-3(4H)-one;7-phenylthio-2H-1,4-benzoxazin-3(4H)-one;4-methyl-7-phenylthio-2H-1,4-benzoxazin-3(4H)-one;4-methyl-7-phenylsulfinyl-2H-1,4-benzoxazin-3(4H)-one; and6-phenylsulfonyl-2H-1,4-benzoxazin-3(4H)-one or a salt, pro-drug orsolvate thereof.
 2. A compound according to claim 1 wherein R₁ is amino,C₁₋₄alkoxy, hydroxy or —OPO₃H₂, wherein the amino group is optionallysubstituted by an amino acid residue and the hydroxy group is optionallyesterified.
 3. A compound according to claim 2 wherein the amino acidresidue is independently derived from: glutamic acid, serine, threonine,arginine, glycine, alanine, β-alanine or lysine.
 4. A compound accordingto any one of claims 1 to 3 wherein X is selected from: —O—, —S— or—N(R₄)—.
 5. A compound according to any one of claims 1 to 4 wherein R₂is hydrogen.
 6. A compound selected from:7-[(2-aminophenyl)sulfanyl]-2H-1,4-benzoxazin-3(4H)-one;7-(3-aminophenoxy)-2H-1,4-benzoxazin-3(4H)-one;6-[2-(4-toluidino)acetyl]-2H-1,4-benzoxazin-3(4H)-one;7-[(3-aminophenyl)sulfanyl]-2H-1,4-benzoxazin-3(4H)-one; and7-[(4-hydroxyphenyl)sulfanyl]-2H-1,4-benzoxazin-3(4H)-one; or salt,pro-drug or solvate thereof.
 7. The use of a compound of Formula (I) asa medicament, wherein:

X is selected from: —O—, —S—, —S(O)—, —S(O₂)—, —N(R₄)—, or—N(R₄)CH₂C(O)—; R₁ is independently selected from: amino, halo, hydroxy,—OPO₃H₂, C₁₋₄alkyl, C₁₋₄alkoxy, N—C₁₋₄alkylamino, N,N-di-C₁₋₄alkylamino,C₁₋₄alkanoylamino or C₁₋₄alkylthio wherein the amino group is optionallysubstituted by an amino acid residue and the hydroxy group is optionallyesterified; R₂ is selected from: hydrogen or C₁₋₄alkyl; R₃ is selectedfrom: hydrogen or C₁₋₄alkyl; R₄ is selected from: hydrogen or C₁₋₄alkyl;n is 0, 1 or 2; and p is 0, 1, 2 or 3; or a pharmaceutically-acceptablesalt, pro-drug or solvate thereof.
 8. A pharmaceutical compositioncomprising a compound according to any one of claims 1 to
 7. 9. The useof a compound of Formula (I) or Formula (II) orpharmaceutically-acceptable salt, pro-drug or solvate thereof, for themanufacture of a medicament to inhibit and/or reverse and/or alleviatesymptoms of angiogenesis and/or any disease state associated withangiogenesis.
 10. The use of a compound of Formula (I) or Formula (II),or pharmaceutically-acceptable salt, pro-drug or solvate thereof, in themanufacture of a medicament for administration in divided doses for usein the production of a vascular damaging effect in a warm-bloodedanimal.
 11. A process for preparing a compound of Formula (I) or Formula(II), or salt, pro-drug or solvate thereof, which process (wherein n, p,X, R₁, R₂, R₃, and R₄ are, unless otherwise specified, as defined inclaim 1), comprises: a) for compounds of Formula (I) or Formula (II)wherein X is —O—, —S— or —N(R₄)—, reacting a compound of Formula (A)with a compound of Formula (B),

 wherein L₁ is a leaving group; b) for compounds of Formula (I) orFormula (II) wherein R₂ is hydrogen, reduction of a compound of Formula(C), wherein R₆ is hydrogen or an alkyl chain,

c) for compounds of Formula (I) or Formula (II) wherein R₂ is C₁₋₄alkyl,reacting a compound of Formula (I) wherein R₂ is hydrogen with asuitable alkylhalide; d) for compounds of Formula (I) or Formula (II)wherein X is —S(O)—, —S(O₂)—, oxidising a compound of Formula (D),

e) for compounds of Formula (I) or Formula (II) wherein X is—N(R₄)CH₂C(O)—, reacting a compound of Formula (E) with a compound ofFormula (F),

 wherein L₂ is a leaving group; and thereafter if necessary: i)converting a compound of Formula (I) or Formula (II) into anothercompound of Formula (I) or Formula (II) respectively; ii) removing anyprotecting groups; iii) forming a salt, pro-drug or solvate.